127 HCC cases, 21 NMLD patients, and 42 health control | Preoperative peripheral blood | 7.5 mL blood stained with immune antibody | Epithelial CTC: PanCK; M-CTC: vimentin, and epithelial–mesenchymal CTC: PanCK, and vimentin | Total CTC number and M-CTC percent were positively correlated with the tumor characteristics, thrombosis, MVI, AJCC stage, BCLC stage, poor RFS rate, and high recurrence risk. | The study focused on EMT-associated CTC subtypes, potentially overlooking CTC heterogeneity. With a limited sample size (n = 127), the conclusions required further validation in larger, independent studies. | 24 |
105 HCC cases | Before radical surgery | 5 mL blood, CanPatrol, and ISH | Epithelial CTCs (EpCAM and CK8/18/19) and M-CTCs (Vimentin and Twist) | M-CTC positivity was significantly higher with AFP, tumor size, multiple tumors, poorly differentiated tumors, incomplete tumor capsule, BCLC stage B or C, MVI, PVTT, and Ki67. | This single-center retrospective study, with a sample size of 105, indicated a risk of selection bias and limited generalizability to other patient populations. The criteria for Ki67 expression grouping should be further validated in additional studies. | 31 |
127 HCC patients | Before surgery | 5mL peripheral blood samples with the CanPatrolTM system and expression levels of target genes in CTCs were assessed using the RNA-ISH method. | Leukocyte biomarker (CD45), epithelial biomarkers (Epithelial cell adhesion molecule, EpCAM; CK8/18/19), and mesenchymal biomarkers (vimentin and Twist) | 1. CTCs served as independent factors associated with the early recurrence in HCC. 2. Survival curve analysis indicated that patients with CTCs experience a shorter recurrence-free survival period. 3. The combination of CTCs and platelet count represented a more robust prognostic marker than their individual use. | This single-center retrospective study with 127 samples needed validation through multi-center prospective studies. CTC detection via filtration may yield false negatives due to tumor cell size variability. As most HCC patients also had HBV, further research is needed to confirm applicability in HCC patients with other liver diseases. | 32 |
56 HCC patients under liver transplantation | Before the operation and seven to ten days after the operation | 5mL peripheral blood samples with CanPatrol CTC-enrichment technique platform and CTCs detected for ISH and fluorescence staining | Epithelial biomarkers (EpCAM and CK8/18/19) and two interstitial biomarkers (Vimentin and Twist) | The postoperative recurrence rates at one, two, and three years for interstitial CTC-positive groups were 21.7%, 37.5%, and 55.5%, respectively, compared to a consistent rate of 10.8% in the CTC-negative group. The recurrence rates at one, two, and three years for the group with increasing interstitial CTCs were 25.2%, 36.9%, and 66.9%, respectively, while they were 12.6%, 24.4%, and 24.4% in the group with decreasing or unchanged CTC. | These changes in total CTC count before and after surgery did not reliably assess liver transplant prognosis in HCC patients, potentially due to differences in inclusion criteria and post-operative immunosuppression protocols. Furthermore, the non-blinded, single-center design and small sample size (n = 56) may introduce bias. | 53 |
73 HCC patients | N.D. | 8 mL blood was detected by immunoaffinity-based method | Epithelial cell adhesion molecule (EpCAM, CD326) and MUC1 | 1. The detection rate of CTCs increased with advancing stages of BCLC. 2. Patients with lower CTC counts exhibited significantly longer overall survival. 3. The combination of CTCs and AFP yielded a higher HR compared to the individual impact of CTCs or AFP alone. | A single-center, prospective cohort study (n = 73) quantified only epithelial CTCs, without mesenchymal or hybrid epithelial-mesenchymal CTCs. Additionally, CTCs were measured at a single time point; serial measurements would offer more information for better outcome prediction. | 25 |
40 HCC patients | N.D. | 4 mL blood | 1. NanoVelcro CTC Assay with round/ovoid cells, DAPI+/CD45−/CK+, with sizes > 6 µm. 2. The NanoString nCounter platform for quantification of selected mRNA transcripts | The HCC-CTC Risk Score panel, which included ten prognostic genes (DDR1, EHHADH, AR, LUM, HSD17B6, PMEPA1, TSKU, NECAB2, LAD1, and SLC27A5), has been validated as an independent predictor of survival. | Patients were recruited from a single institution (n = 40), with a limited representation of different stages, tumors, and treatment characteristics. The analysis of gene expression in CTCs was a proof-of-concept study; further genomic refinement could enhance the prognostic power and utility of the relevant genes. | 64 |
193 HCC patients | Pre- and post-operative | 5 mL peripheral blood | ChimeraX ® -i120 CTC detection platform with epithelial cell (EpCAM), (Campos-Murguia, #225), CK19, and single-cell whole genome sequencing. | 1. HCC patients experienced a reduction in the burden of CTCs following liver transplantation. 2. CTCs served as post-transplantation biomarkers in HCC patients, aiding in the assessment of recurrence risk. | The small cohort size (n = 193), short follow-up, single-center design, and disease heterogeneity between groups reduced the predictive value of preoperative CTC counts for tumor recurrence. Additionally, CTCs were only measured at two post-transplant time points, insufficiently assessing the predictive value of continuous CTC monitoring for recurrence. | 52 |
160 HCC patients | Before surgery | 15 mL of peripheral blood sample | CanPatrol™ CTC enrichment technology with RNA-ISH, Cancer stem cells marker (Nanog), epithelial marker (CK8, 18 and 19 and EpCAM), and mesenchymal marker (Twist and Vimentin) | 1. The quantity of CTC expressing RNA for both EpCAM and Nanog was closely associated with postoperative recurrence of HCC. 2. Patients with CTCs showing higher expression of Nanog exhibited a higher recurrence rate. 3. CTC expressing Nanog was predominantly categorized into the mixed CTC and M-CTC subtypes. 4. The number of CTCs and the expression of Nanog were significantly correlated with BCLC stage, vascular invasion, tumor size, and HBV. | A single-center, prospective study (n = 160) collected CTCs only preoperatively, lacking post-surgery follow-up. Consequently, Nanog gene expression could only be correlated with preoperative CTC subtypes. | 46 |
7 healthy donors, 14 liver cirrhosis patients, and 31 HCC patients | ND | 3 mL peripheral blood samples | MCA system for size-based isolation of CTCs and immunofluorescence staining with positive for CK and DAPI | The positivity rate of CTCs in HCC was significantly higher than that in LC, leading to a reduction in the cumulative survival rate of HCC patients, particularly those with localized HCC. The quantity of CTCs in metastatic HCC was significantly higher than in localized HCC. The expression of AFP, GPC3, EpCAM, and ALB genes was detected in isolated CTC, with the detection rate of ALB mRNA significantly higher in the metastatic group compared to the localized group. | This study focused on MCA analysis of a small number of clinical samples. Analyzing more samples would have better evaluated the system’s applicability to CTC counting and tumor characteristics. Additionally, the MCA system’s reliance on specific antibodies limited its broader use. | 44 |
204 HCC patients | Before surgery | 7.5 mL peripheral blood | CellSearch™ System with CK positive for CTC detection. CTC clusters were detected as an aggregation of CTCs containing two or more distinct nuclei and with contiguous cytoplasmic membranes | 1. In preoperative samples, 37.3% of patients were detected with CTCs, and 9.3% with CTC clusters. The survival period of patients with CTCs ≥ 2 was significantly shorter than that of patients with CTC < 2. 2. Patients with CTC clusters had a significantly poorer prognosis, and their survival time was also significantly shorter than those without CTC clusters. The presence of CTC clusters in HCC was associated with the activation of the Wnt/β-catenin signaling pathway. | The low detection rate and quantity of CTC clusters and CTCs in this study may have been attributed to the use of the CellSearch system, potentially leading to an underestimation of cluster numbers. Most CTCs within clusters were EpCAM-positive without EMT characteristics, which differed from other studies. This study focused only on the tumor cell component within CTC clusters, without analyzing the non-tumor cell elements. | 63 |
41 HCC patients (31 patients underwent liver transplantation/10 patients underwent surgical resection) | Before surgery, at post-operative day 5 and at day 30 | 7 mL peripheral blood samples | The Isoflux® system (Fluxion Biosciences) | In HCC patients undergoing LR, CTC clusters were found more frequently 30 days postoperatively compared to patients undergoing LT. This difference arose from the incomplete clearance of CTCs and negatively impacted survival. | The prospective cohort study had a limited sample size due to slow accrual, extended monitoring, and increased CTC analysis costs. Additionally, the study only examined CTC clearance kinetics within the first month post-surgery, without long-term insights into CTC-immune system interactions. | 56 |
17 HCC patients (10 patients underwent microwave ablation and 7 patients underwent conventional transarterial chemoembolization) | Before and after radiological interventions | 10 mL peripheral blood samples | Flow cytometry with ASGPR, CD146 and CD274 (PD-L1) | The rate of CTCs in HCC patients was significantly decreased after MWA treatment. | The prospective single-center study had a small patient cohort, and the heterogeneity and rarity of CTC antigens made detection and analysis challenging. | 51 |
179 HCC patients | ND | 5 mL peripheral blood samples | The CanPatrol CTC enrichment technique with RNA-ISH with epithelial cell markers (EpCAM and CK8/18/19), and mesenchymal cell markers (vimentin and Twist). Survivin expression in CTCs was assessed using the RNA-ISH method. | The counts of CTCs and survivin-positive CTCs were significantly higher in the HCC patients and associated with tumor stage and differentiation degree. | The single-center case study had limited follow-up, preventing analysis of the relationship between survivin-positive CTC counts and overall survival. | 26 |
214 HCC patients | Before surgery | 7.5 mL peripheral blood samples | CanPatrol™ CTC analysis technology, epithelial cells were labeled with EpCAM and CK8/18/19, and M-cells were labeled with vimentin/twist. Epithelial CTC, M-CTC, and mixed CTC subtypes can form CTC-WBC clusters with WBCs. | Both CTC clusters and the total number of CTCs associated with tumor size and number, portal vein tumor thrombus, BCLC stage, and AFP level. CTC clusters in HCC patients were an independent prognostic indicator of DFS and OS. | The single-center retrospective cohort study cannot be generalized to non-Asian populations. It primarily focused on resectable HCC patients, lacking data on advanced-stage liver cancer cases. | 27 |
136 HCC patients | Before resection | 5 mL peripheral blood samples | The CanPatrol system with RNA-ISH EpCAM, CK8/18/19 (epithelial biomarkers), and vimentin and twist (mesenchymal biomarkers). | In patients with a lower quantity of CTCs and a negative mesenchymal and epithelial/M-CTC phenotype, tumor-free survival rates were significantly higher. Higher pre-resection CTC counts and positive mesenchymal and epithelial/M-CTC phenotypes were significantly associated with extrahepatic and multi-intrahepatic recurrence. | This single-center retrospective study used the CanPatrol system, which only collected CTCs larger than 8 µm, potentially missing smaller CTCs. | 38 |
344 HCC patients | Preoperative | 7.5 mL peripheral blood samples | CellSearch system with positive for CK8/18/19 and/or EpCAM | Patients with CTCs undergoing TACE demonstrated significantly enhanced clinical outcomes and a substantial reduction in early recurrence. | This single-center retrospective study did not evaluate the value of post-operative CTCs in guiding adjuvant TACE, and PSM analysis did not account for confounding factors or bias. Additionally, the CellSearch system did not fully assess CTC status. | 39 |
50 HCC patients | Before liver transplantation | 3.2 mL peripheral blood | imFISH, which combined the FISH probes with chromosome 8 (orange) centromere probes CTC cells positive with CEP8+/DAPI+/CD45− | The CTC number was correlated with tumor size, AFP level, tumor grade, and Recurrence. CTC-negative patients had higher one-year disease-free survival rates. | This single-center prospective cohort study collected only preoperative CTCs, limiting the ability to assess the relationship between post-operative CTCs and early recurrence after LT. Extending the follow-up to three to five years would be more meaningful. | 47 |
217 HCC patients | ND | 7.5 mL peripheral blood | Ficoll solution was incubated with fluorescent antibodies including CTC positive (EpCAM; panCK19), and isolated by flow cytometry. | USP1 was frequently upregulated in CTCs and correlated with metastasis and reduced overall survival rate in HCC patients. | This single-center case study did not investigate real-time changes in patient CTCs. While it aimed to show that immune attacks limit CTC survival, the evidence was insufficient to establish the association between USP1 and immune evasion. | 60 |
99 HCC patients | Pre-operatively | 5 mL peripheral blood samples | CanPatrolTM with epithelial biomarker probes (EpCAM and CK8/18/19), interstitial biomarker probes (vimentin and twist) | Positive CTCs exhibited positive correlations with BCLC staging, tumor diameter and quantity, capsule integrity, MVI, portal vein thrombosis, AFP, and hepatitis B DNA in HCC. Higher expression of CXCR4 was more commonly observed in mixed CTCs than mesenchymal CTCs. | This single-center prospective analysis did not fully establish the link between CTCs and AFP. Only 22 patients provided survival-related follow-up data, and overall survival was not included. | 33 |
137 HCC patients | Before surgery, during surgery (30 m after tumor removal) and 1 week, 1/2/3/6 months, and 1 year after surgery | 5 mL blood samples | Isolation by Size of Epithelial Tumor Cells | 1. Preoperative CTC count was an independent predictor of MVI. 2. CTC count had better predictive value than AFP and tumor diameter. 3. The number of CTCs in the non-early recurrence group decreased significantly after surgery 4. Patients with CTC count ≥ 5 had worse long-term outcomes. | This single-center case study had a short follow-up period. The ISET method may have caused non-CTC to clog filter pores and lead to background contamination, resulting in failed CTC isolation in some samples. | 34 |
87 HCC patients (49 early-stage, 22 locally advanced, and 16 metastatic), 7 cirrhosis patients, and 8 healthy controls | ND | 4 mL blood samples | CTCs were separated by gradient centrifugation with Ficoll-Paque solution then the NanoVelcro Chip defined CK+ CTCs were defined as round/ovoid cells (DAPI+/CK+/PD-L1−/CD45−) and PD-L1+ CTC are the subpopulation of HCC CK+ CTC defined as round/ovoid events (DAPI+/CK+/PD-L1+/CD45−) | PD-L1+ CTCs were predominantly found at advanced stage and had a significantly worse OS in HCC patients. | This single-center case study required additional testing for EMT markers beyond EpCAM to identify CTC surface antigens. The association between PD-L1+ CTCs and immunotherapy needed further validation. | 61 |
197 HCC patients (retrospective training cohort (144 patients) or a prospective validation cohort (53 patients)) | Before surgery and one month after surgery | 7.5 mL peripheral blood | CELLSEARCH system | The presence of EHM was significantly correlated with a higher postoperative CTC burden. Patients with a postoperative CTC count ≥ 3 faced an elevated risk of EHM and a shorter median overall survival. | This single-center case study collected only EpCAM+ CTCs, missing other heterogeneous CTCs, which reduced the sensitivity and specificity of CTC detection. | 54 |
309 HCC patients | Before surgery | 7.5 mL peripheral blood | CellSearch and positive CTC was defined as CTC ≥ 1 | CTC-positive HCC patients exhibited higher MVI and greater FMT. Within the CTC-positive group, a surgical margin > 1 cm independently provided protection against early recurrence and was associated with a lower early recurrence rate. | The single-center retrospective study could not eliminate potential selection bias or confounding factors. Most enrolled patients were infected with HBV, and applicability to HCC from other causes needs further validation. Additionally, only EpCAM+ CTCs were collected, missing other heterogeneous CTCs. | 35 |
85 HCC patients | Before surgery | 8 mL peripheral blood | Immuno-magnetic positive enrichment (GPC3) coupled with flow cytometry (CK7/8) | Patients had higher GPC3-positive CTC counts with a higher incidence of mPVI, a lower disease-free survival, and a lower overall survival. | This single-center case study did not analyze liver transplantation or adjuvant therapy. Additionally, flow cytometry for CTCs may have included non-specific events. | 62 |
105 HCC patients and 132 control | Before and after surgery | 5 mL peripheral blood | A tapered slit filter platform based on the cell size and morphology with positive for 4′,6-diamidino-2-phenylindole, and CK | After surgery, HCC patients exhibiting an increase in CTCs were significantly associated with higher recurrence rates. CTC counts were considered as an independent predictive factor for predicting PFS. Patients with low alpha-fetoprotein levels and cirrhosis, along with positive CTCs, were correlated with lower survival rates and higher recurrence rates. | This single-center prospective cohort study might not isolate smaller or highly deformable CTCs through the TSF platform, limiting the sensitivity and specificity. | 55 |
42 HCC patients and 5 control | ND | 10 mL peripheral blood | Labyrinth microfluidic device | 1. CTC positive rate was elevated in advanced HCC stages. 2. In 71.4% of HCC patients, the cancer stem cell marker CD44 was exhibited in CTCs. 3. CTM was present in 55% of HCC patients and was associated with advanced HCC stages. | This single-center case study required further research to determine the association between tumor invasion and CD44+ CTCs or CTMs. | 28 |
176 HCC patients | Before chemotherapy or radiotherapy | 5 mL of peripheral blood | CanPatrol with multiplex RNA with ISH; epithelial (CK8/18/19, and EpCAM) and mesenchymal (Vimentin and Twist) | 1. All types of CTCs were more numerous in HCC patients. 2. BCLC stage B-C had more M-CTCs than BCLC stage 0-A. | Due to the small subgroup size in this single-center prospective cohort study, the association between total CTC count and other factors could not be effectively analyzed. | 29 |
126 HCC patients | Before and after cancer treatments | 7.5 mL peripheral blood | Immuno-magnetic beads and Immunostaining-fluorescence ISH (EpCAM, CK18, PD-L1, and Vimentin) | CTC count was higher in HCC stages III and IV than in stages I and II. | The single-center case study lacked follow-up and prognostic assessment of patients. Performing multiploidy analysis on different chromosomes in CTCs could help clarify the relationship between multiploidy and cancer stages. | 30 |
62 HCC patients | Postoperative | 5 mL of peripheral blood | CanPatrol with multiplex RNA-ISH; epithelial (CK8/18/19; EpCAM) and mesenchymal (Vimentin and Twist). | M-CTCs were associated with shortened postoperative disease-free survival and acted as independent risk factors for early recurrence. | The single-center prospective cohort study should further examine the impact of different surgical methods on CTC phenotypes and counts. The extended follow-up aimed to clarify the link between CTC subtypes and patient survival. | 49 |
325 HCC patients, 201 chronic hepatitis B infection and liver cirrhosis patients, 100 benign hepatic lesion patients, and health 260. | Before initial diagnosis or one week after tumor resection | 5 mL of peripheral blood | RosetteSep Human CD45 Depletion Cocktail and mRNA expression levels of 10 target genes (EpCAM, CD133, CD90, CK19, ABCG2, CD24, CD44, ICAM1, Nestin, and β-actin) | 1. A multiple-marker CTC detection panel distinguished early and AFP-negative HCC from CHB, LC, and BHL. 2. CTC load decreased after tumor resection, and patients with a high CTC load were associated with tumor recurrence postoperatively. | The multi-center prospective cohort study included HCC patients with cirrhosis or HBV. Validation in other regions and HCC types was needed. | 20 |
43 HCC patients with HCC | Before and after percutaneous radiofrequency ablation | 5 mL of peripheral blood | CanPatrol with ISH; epithelial (EpCAM and CK8/18/19); mesenchymal (vimentin and twist) | The levels of M-CTCs increased and were associated with a decrease in lymphocyte count following hepatic tumor PRFA. | The single-center prospective cohort study used RNA-ISH for CTC detection, differing from antigen-antibody methods. The correlation between CTC levels and immune cell subsets (CD3+8+ T cells and NK cells) was not well established. | 48 |
139 HCC patients and 23 control | Before and after operation | 7.5 mL peripheral blood | CellSearch with EpCAM and CK | 1. Postoperative CTC counts increased, associated with the macroscopic tumor thrombus status, and were correlated with worse disease-free and overall survival rates. 2. Patients with preoperative high CTC counts had poor prognoses. | The single-center prospective cohort study only detected the EpCAM CTCs. | 37 |
47 HCC patients | After liver transplantation | 5mL of blood samples | CanPatrol with ISH; epithelial (EpCAM and CK8/18/19); mesenchymal (vimentin and twist) | The levels of epithelial and interstitial CTCs increased following LTx and continued to rise over the follow-up period. | The single-center case study, limited by sample size and the collection of post-transplant samples only for some patients, could not fully compare the association between CTCs and HCC recurrence after liver transplant, nor the differences in CTCs before and after transplant. | 57 |
112 HCC patients | Before and after resection | 5 mL peripheral blood | CanPatrol with RNA-ISH, epithelial (EpCAM, CK8/18/19), and mesenchymal (vimentin and Twist) | 1. CTC count ≥ 16 and M-CTC percentage ≥ 2% were associated with early recurrence, multi-site intrahepatic recurrence, and lung metastasis before resection surgery. 2. Increased CTC count and M-CTC percentage were associated with recurrence after surgery. 3. The overexpression of BCAT1 in CTCs may trigger the EMT process, inducing CTC release. | The single-center prospective cohort study using the CanPatrol system may miss smaller CTCs and only examined the relationship between BCAT1 expression and CTCs. | 19 |
73 HCC patients | Before resection | 7.5 mL blood | CellSearch EpCAM-positive (panCK8/18/19) and Microfluidic qRT-PCR | 1. The total CTC count from hepatic veins was correlated with the initiation of EMT in HCC. 2. The burden of CTCs and circulating tumor microemboli from HV could predict intrahepatic recurrence and postoperative lung metastasis. 3. CTCs were primarily epithelial upon release but switched to an EMT phenotype during dissemination through the bloodstream, involving associated Smad2 and β-catenin protein signaling pathways. | The single-center prospective cohort study limited the prognostic significance of CTCs in different vascular regions. The molecular mechanisms behind CTC spatial heterogeneity needed further investigation. | 23 |
61 HCC patients, 11 had cirrhosis without HCC, adenoma, focal NMLD, and eight healthy controls | Before and after surgery | 4 mL blood | NanoVelcro CTC Assay with ASGPR, GPC3, EpCAM, and Vimentin. HCC CTCs were defined as round/ovoid cells, DAPI+/CD45–/CK+, with a size ≥ 6 µm. EMT phenotype, VIM-positive CTCs are the subpopulation of HCC CTCs defined as round/ovoid events, DAPI+/CD45–/CK+/VIM+, with size ≥ 6 µm | VIM-positive CTCs not only predicted OS of HCC patients but also accurately distinguished patients in the early stage, LT eligible, from those in the locally advanced/metastatic stage, LT ineligible. Furthermore, VIM-positive CTCs indicated a faster recurrence trend after surgical or local treatment of potentially curable early-stage HCC. | The single-center prospective cohort study used EpCAM, ASGPR, and GPC-3 to isolate CTCs but lacked EMT markers. Additionally, it analyzed vimentin(+)-CTCs without considering other CTC markers. | 59 |
42 HCC patients | Before and after surgery | 5 mL peripheral blood | CanPatrol with RNA-ISH | 1. CTCs were associated with the Edmondson stage in HBV-related HCC before surgery. 2. Postoperative CTC counts and the change in CTC counts before and after surgery were associated with PFS. 3. The postoperative CTC count was associated with TP53 expression. | This single-center prospective cohort study only assessed total CTC counts, lacking further analysis of CTC phenotypes and genotypes. | 45 |
63 HCC patients, 31 chronic liver disease patients, and 26 health control | Before or after surgery | 5–15 mL of blood | Microfluidic CTC-iChip | CTCs were significantly detected in 56% of untreated HCC patients compared to 3% of nonmalignant liver disease patients, indicating a risk for HCC. | The single-center case study required establishing screening criteria and validating the sensitivity and specificity of RNA-based CTC detection. | 17 |
57 HCC patients | Before surgical treatment | 7.5 mL blood | CellSearch with EpCAM and CK8/18/19 | CTC-positive patients had a significantly higher risk of recurrence with a HR of 2.3, and a shorter RFS. | This single-center prospective study did not examine EMT markers in CTCs, which may have reduced the number of detected CTCs and limited the ability to link CTCs to HCC recurrence. | 40 |
123 patients (HCC = 52) and 12 normal controls. | ND | 5 mL blood samples | Flow cytometry analysis with the EpCAM. | High karyoplasmic ratios were more prevalent in HCC patients with MVI than in both HCC and non-cancer patients. | This single-center study used flow cytometry to identify high karyoplasmic ratio cells as CTCs without validation standards, risking misidentification. Additional markers are needed for verification. | 36 |
49 HCC patients | Before operation | 6 mL blood | EpCAM mRNA+ CTCs | 1. The counts of EpCAM mRNA+ CTCs and Treg/CD4+ cells showed a significant correlation with postoperative HCC recurrence. 2. High CTC/Treg level indicated a higher risk of postoperative HCC recurrence, significantly increasing the one-year recurrence rate. | The single-center prospective cohort study included only HBV-induced and early-stage HCC patients. The relationship between CTCs and Treg cells required further analysis. | 41 |
72 patients | After hepatectomy at zero, three, six, nine and twelve months | 3 mL blood samples | Anti-EpCAM nanoparticals with magnet | 1. The positive expression of AFP mRNA in CTCs could serve as a predictor for metastasis both before and after hepatectomy. 2. The release of AFP expression by HCC into circulation was inevitably a primary source of HCC metastasis. | This single-center cohort study used only EpCAM to detect CTCs, while AFP nested RT-PCR limited technical issues, causing discrepancies. | 58 |
69 patient and 31 control samples (15 healthy volunteers and 16 patients with cirrhosis without cancer) | ND | 12 mL blood samples | Immunofluorescence of panCK4/5/6/8/10/13/18, EpCAM, AFP, GPC3, and DNA-PK | 1. CTC number associated with tumor size and PVT. 2. HCC patients with fewer than one CTC had a median survival period exceeding 34 months, compared to patients with more than one CTC, whose median survival was only 7.5 months. | The single-center cohort study showed a weak association between peripheral neutrophil count and portal vein invasion. | 42 |
20 HCC and 10 NMLD patients | Prior biopsy or resection | 7.5 mL blood samples | CellSearch by immunomagnetic EpCAM enrichment and fluorescence-activated cell sorting | 1. The CTC counts were associated with AFP levels and vascular invasion. 2. The frequency of low-variant DNA was higher in CTCs. | This single-center case-control study showed a limited match between CTCs, FFPE tumors, and PBMC DNA, with CTCs having lower WGA coverage than FFPE. | 22 |
40 liver cancer patients and 27 healthy volunteers | Before surgery or treatment | 5 mL blood samples | CanPatrol CTC enrichment technique with RNA-ISH | 1. Three subtypes of CTCs were identified using EMT markers, including epithelial CTCs, epithelial/M-CTCs, and M-CTCs. It was observed that M-CTCs were more prevalent in patients with cancer metastasis. | The single-center case-control study isolated CTCs using filtration and RNA-ISH, a more complex technique compared to immunostaining, with non-standardized probes. | 4 |
26 HCC patients | Preoperative, postoperative, and 24 h after surgery | 10 mL blood samples | Quantitative flow cytometry with CD45−/CD44+/CD90+ cells | HCC patients undergoing laparoscopic surgery exhibited fewer CTCs and lower secretion of IL-6 and IL-8 compared to those undergoing traditional open surgery. | This single-center prospective cohort study found that the accuracy of CTC detection needed improvement. Patient variability also affected CTC detection, and further analysis was required to explore the relationship between cytokines and CTCs. | 50 |
299 HCC patients (Resection, n = 157; TACE, n = 76; radiotherapy, n = 66) | Pretreatment and post-treatment | 7.5 mL of blood | Negative enrichment and qRT-PCR-based CTC detection platform and CellSearch system with EpCAM+ CTC | The preoperative levels of CTCs could predict the prognosis of patients with HCC undergoing liver resection, TACE, and radiotherapy. Moreover, an increase in CTC levels after treatment indicated disease progression. | The single-center prospective cohort study had a short follow-up period, and qRT-PCR for EpCAM was prone to leukocyte contamination, leading to a higher false positive rate. | 21 |
11 HCC patients | During treatment | 20 mL of blood | CTC type detection by immunofluorescence staining with epithelial panCK and mesenchymal markers (vimentin, N-cadherin). | Different CTC subtypes could be identified in the peripheral blood of HCC patients. Changes in the ratio of epithelial to M-CTCs were associated with a longer median time to progression. | The single-center cohort study observed that CD45-positive cell depletion during negative enrichment led to CTC loss. Additionally, immunomagnetic methods showed lower recovery rates compared to density gradient centrifugation. | 43 |
60 HCC patients, 10 patients with benign liver diseases (including patients with cirrhosis, chronic hepatitis B, hepatic hemangioma, and liver cysts), 10 healthy volunteers and 10 patients with miscellaneous advanced cancers other than HCC | ND | 10 mL of blood | ASGPRs with immunofluorescence staining | In HCC tumors, the presence of positive CTCs was associated with the expression levels of E-cadherin, vimentin, and twist. The expression of E-cadherin indicated a significant role for EMT in facilitating the blood-borne dissemination of primary HCC cells. Moreover, the expression levels of twist and vimentin in CTCs could serve as biomarkers for evaluating metastasis and prognosis in HCC patients. | The single-center study used ASGPR, a receptor expressed only in HCC, to detect EMT CTCs. | 18 |
85 HCC patients, 37 patients with benign liver diseases, 20 healthy volunteers, and 14 patients with other cancers. | ND | 5 mL blood | HCC cells were bound by biotinylated asialofetuin and magnetically labeled | CTCs were identified in 81% of HCC patients. The positivity rate and quantity of CTCs were significantly correlated with tumor size, portal vein tumor thrombosis, differentiation status, as well as TNM classification and Milan criteria-based disease staging. Additionally, HER-2 gene amplification and TP53 gene deletion were detected in CTCs. | The single-center cohort study used ASGPR and Hep Par 1 techniques for CTC detection, which were applicable only to HCC, while HER-2 amplification in CTCs required further investigation. | 15 |
44 HCC patients, 30 patients with chronic active hepatitis, 39 with liver cirrhosis, and 38 healthy individuals | ND | 6 mL peripheral blood | The ISET method with filtration and β-Catenin mutations assay | CTCs were associated with tumor diffusion, portal vein tumor thrombosis, and shorter overall survival. | The single-center cohort study using the ISET method for detecting CTCs may have missed those with diameters less than 25 µm | 16 |