[Show abstract][Hide abstract] ABSTRACT: Background:
The distinct expression pattern of tumour-associated antigens (TAAs) might be a critical reason for the inefficacy of immunity-based treatments and heterogeneous postsurgical recovery in patients with solid tumours, including hepatocellular carcinoma (HCC). However, little is known about the clinical value of the coexpression patterns of multiple TAAs.
We determined the expression of multiple TAAs with identified immunogenicity (GPC3, AFP, SSX-2, NY-ESO-1, EpCAM, midkine) and the density of tumour-infiltrating immune cells by immunohistochemistry in a panel of 362 primary HCC patients. We evaluated the association between the TAAs, immune cell infiltration, clinicopathological parameters, and prognosis.
Patients who coexpressed more TAAs had better prognosis (P<0.00001, overall survival). The integrated pattern of TAA was associated with good differentiation and small tumour size, and with more CD57+ natural killer and CD20+ B-cell infiltration (P<0.05). Multivariate Cox proportional hazards analysis identified the TAA index as an independent prognostic indicator (hazard ratio 0.625; 95% confidence interval 0.467–0.837; P=0.002), and could further predict patient prognosis in collaboration with local immune infiltration.
Our results could provide new evidence for the improvement of prognostic molecular signatures in HCC, and a novel rationale for patient enrolment in future immunotherapeutic trials and/or clinical treatments.
British Journal of Cancer 07/2013; 109(4). DOI:10.1038/bjc.2013.390 · 4.82 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To develop an in situ molecular signature to predict postsurgical recurrence in hepatocellular carcinoma (HCC) patients.
Immunohistochemistry was performed using tissue microarrays containing both tumoral and peri-tumoral regions of the advancing tumor edge from 336 HCC patients (289 were positive for hepatitis B virus) who underwent curative resection. Forty-nine variables were analyzed in the training set (n=151) using support vector machine and stepwise algorithms to develop a classifier to predict recurrence within 1 year, which was mainly caused by invasion or metastasis from the primary tumors. The classifier was further validated in an independent cohort of 185 patients (71 internal and 114 external).
The final signature was composed of eight IHC features: CD80(T), B7-DC(T), HLA-DR(P), FasL(P), Bcl-2(T), Ki-67(T), cyclin D1(T), and CK19(T). In the independent test set, this classifier reliably predicted recurrence within 1 year (sensitivity, 69.1%; specificity, 65.0%) with an odds ratio of 4.149 (95% CI, 2.189-7.864). Based on a multivariate logistic model, the in situ molecular signature provided significant predictive power independent of tumor number, tumor size, vascular invasion and BCLC classification (p=0.001). The highest potential clinical impact of the classifier was observed in early-stage (BCLC classification 0-A) patients (p<0.0001), and the classifier was also predictive of the time-to-recurrence and overall survival (both p<0.0001).
This in situ molecular classifier could provide a novel approach to identify patients who are at greatest risk for postsurgical recurrence of HCC and may benefit from intensive clinical follow-up or chemopreventive strategies.
Journal of Hepatology 04/2012; 57(2):313-21. DOI:10.1016/j.jhep.2012.03.027 · 10.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Distinct morphologic features of microvascular endothelium exist in tumor tissues. The objective of this study was to investigate the prognostic value of endothelium-coated tumor clusters (ECTCs) in hepatocellular carcinoma (HCC).
ECTCs were evaluated by immunohistochemical staining for cluster of differentiation 34 (CD34) (a cell surface glycoprotein which is expressed specifically on tumor microvascular endothelium in HCC) in 239 specimens from patients with primary HCC. Overall survival (OS) and time to recurrence (TTR) were determined using Kaplan-Meier analysis and a Cox proportional hazards regression model. Levels of terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, and Ki-67 expression, and E-cadherin expression were assessed to determine tumor apoptosis, proliferation, and invasiveness, respectively.
The presence of ECTCs was associated with a poor prognosis in all patients and in patient subgroups stratified by tumor size, TNM classification, and Barcelona Clinic Liver Cancer stage and tumor invasiveness. In a multivariate Cox proportional hazards analysis, the presence of ECTCs emerged as an independent prognostic indicator of both poor OS (P = .001; hazard ratio, 1.949) and shorter TTR (P < .001; hazard ratio, 2.085). Furthermore, the presence of ECTCs was associated with micrometastatic endothelium-coated emboli (P < .001; chi-square test) and early relapse after resection (P < .001; chi-square test). In addition, patients who had endothelium-coated emboli, in which tumor cells displayed high proliferation and low apoptosis, had poor OS and shorter TTR.
The current results suggested that the presence of ECTCs was an efficient, simple, and convenient predictor of a poor prognosis in patients with HCC that potentially may serve as a novel target for the prevention and treatment of HCC metastasis.
Cancer 11/2011; 117(21):4878-89. DOI:10.1002/cncr.26137 · 4.90 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Macrophages (Mφ) and regulatory T cells (Tregs) are the major components of the inflammatory infiltrate in virtually all tumors. The objective of this study was to investigate the prognostic significance of Mφ and Tregs infiltration in advanced gastric cancer after radical resection.
CD68(+) Mφ and FOXP3(+) Tregs were assessed by immunohistochemistry in tissues from 107 patients with surgically advanced gastric cancer. The microlocalization of Mφ and Tregs cells with respect to the development of gastric cancer were given special concern. Prognostic value of normal, peritumoral, and intratumoral Mφ and Tregs densities was evaluated by Kaplan-Meier analysis and Cox regression.
The results showed that the presence of intratumoral CD68(+) Mφ was an independent prognostic factor for overall survival (OS) (P = 0.02). Moreover, the combination of high numbers of intratumoral CD68(+) Mφ and FOXP3(+) Tregs was associated with improved survival (P = 0.041). Five-year OS rate was only 27% for patients with low intratumoral Mφ and intratumoral Tregs compared with 62% for patients with high intratumoral Mφ and intratumoral Tregs. In addition, advanced intestinal-type gastric cancers were more likely to have fewer infiltrating Mφ than diffuse-type cancers (P = 0.024).
Association of intratumoral Mφ and Tregs is a promising independent predictor for survival in advanced gastric cancer. The results suggested that a combination of concomitant stimulation of intratumoral Mφ and Tregs may be an effective strategy for treatment of patients with advanced gastric cancer after radical resection.
[Show abstract][Hide abstract] ABSTRACT: Macrophages (Mphi) are prominent components of solid tumors and exhibit distinct phenotypes in different microenvironments. We have recently found that tumors can alter the normal developmental process of Mphi to trigger transient activation of monocytes, but the underlying regulatory mechanisms are incompletely understood. Here, we showed that the protein expression of transcription factor C/EBPbeta was markedly elevated in tumor-associated Mphi both in vitro and human tumors in situ. The expression of C/EBP protein correlated with cytokine production in tumor-activated monocytes. Moreover, we found that C/EBPbeta expression was regulated at the post-transcriptional level and correlated with sustained reduction of microRNA-155 (miR-155) in tumor-activated monocytes. Bioinformatic analysis revealed that C/EBPbeta is a potential target of miR-155 and luciferase assay confirmed that C/EBPbeta translation is suppressed by miR-155 through interaction with the 3'UTR of C/EBPbeta mRNA. Further analysis showed that induction of miR-155 suppressed C/EBPbeta protein expression as well as cytokine production in tumor-activated monocytes, an effect which could be mimicked by silencing of C/EBPbeta. These results indicate that tumor environment causes a sustained reduction of miR-155 in monocytes/Mphi, which in turn regulates the functional activities of monocytes/Mphi by releasing the translational inhibition of transcription factor C/EBPbeta.
[Show abstract][Hide abstract] ABSTRACT: Immunosuppression mediated by regulatory T cells (Tregs) is a key facilitator of tumor immune evasion, but the source of these Tregs and their contribution to human cancer progression remains unclear. This study investigated the properties of FoxP3(+) Tregs, their prognostic value in patients with hepatocellular carcinoma (HCC) and the underlying mechanisms of FoxP3(+) Treg intratumoral accumulation. In addition to an increased number of circulating FoxP3(+) Tregs, the results also showed that FoxP3(+) Tregs gathered in the tumor site, where they suppressed tissue-derived CD4(+)CD25(-) T-cell activation (p < 0.001), promoting disease progression and poor prognosis in HCC patients (< 0.01). The intratumoral prevalence of FoxP3(+) Tregs was associated with a high density of macrophages (Mvarphi) (p < 0.001). Depletion of tissue Mvarphi thus attenuated the increase of liver FoxP3(+) Treg frequency attributed to in vivo inoculation with hepatoma (p = 0.01), whereas Mvarphi exposed to tumor culture supernatants from hepatoma-derived cell lines increased FoxP3(+) Treg frequency in vitro (p < 0.001). This increase was partially blocked by antiinterleukin-10 antibody (p < 0.01). In conclusion, tumor-associated Mvarphi may trigger a rise of the intratumoral FoxP3(+) Treg population, which in turn may promote HCC progression.
International Journal of Cancer 10/2009; 125(7):1640-8. DOI:10.1002/ijc.24556 · 5.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Macrophages constitute a major component of the leukocyte infiltrate of tumors and perform distinct roles in different tumor microenvironments. This study attempted to investigate the prognostic values of tumor-infiltrating macrophages in patients with hepatocellular carcinoma after resection, paying particular attention to their tissue microlocalization. The CD68(+) macrophages were assessed by immunohistochemistry in tissues from 137 patients with hepatocellular carcinoma. Prognostic value of intratumoral, marginal, and peritumoral macrophage densities was evaluated by Kaplan-Meier analysis and Cox regression. Both intratumoral and marginal macrophage densities were associated inversely with overall survival (P = .034 and .004, respectively) and disease-free survival (P = .006 and .008, respectively). In contrast, peritumoral macrophage density was associated with neither overall survival nor disease-free survival. Intratumoral macrophage density emerged as an independent prognosticator of overall survival (hazard ratio = 1.721, P = .049) and disease-free survival (hazard ratio = 2.165, P = .007). Marginal macrophage density, but not intratumoral macrophage density, was associated with vascular invasion, tumor multiplicity, and fibrous capsule formation. Our results demonstrate that high macrophage infiltration predicts poor prognosis in patients with hepatocellular carcinoma. These results, together with our previous report showing the distinct activation patterns of macrophages in different areas of tumor tissue, implies that macrophages in those areas may use different strategies to promote the tumor progression.
Human pathology 12/2008; 40(3):381-9. DOI:10.1016/j.humpath.2008.08.011 · 2.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the clinical significance of the amount of regulatory T cells (Treg) in the peripheral blood CD4+ cells and tumor tissue in primary hepatocellular carcinoma (HCC).
From January 1999 to December 2000, 63 HCC patients underwent radical resection in Sun Yatsen University Cancer Center. Tregs in those patients were detected in the samples of preoperative peripheral blood by flow cytometry and also in tissue samples of the resected tumors by immunohistochemistry. All patients had been followed up till Dec 30, 2005. The correlations of Treg amount in the peripheral blood CD4+ cells and tumor tissue with clinicopathologic characteristics and prognosis of HCC were analyzed.
The proportion of Treg/CD4+ in the peripheral blood was significantly higher in the patients with HCC than that in those with HBsAg positive (P < 0.01) and in the normal controls (P < 0.01). The mean number of Treg in tumor tissue was (15.69 +/- 13.29)/mm2, but none or very few Treg was detected in the normal liver tissue, para-cancerous liver tissue, and HBV-infected liver tissue. The proportion of Treg/ CD4+ in the peripheral blood was significantly positively correlated with the number of Treg in tumor tissue (P = 0.024). The 5-year survival in patients with high amount of Treg in both peripheral blood and tumor tissue was significantly poorer than that in the patients with low amount of Treg (P = 0.042, 0.019). The 5-year disease-free survival rate was significantly lower in the patients with high amount of Treg in tumor tissue than that in the patients with lower amount (P = 0.001).
Regulatory T cells in the circulatory blood and tumor tissue are increased in patients with hepatocellular carcinoma. The increased amount of regulatory T cells either in peripheral blood or in the tumor tissue is pertaining to poor prognosis. Detection of regulatory T cells both in the preoperative peripheral blood CD4+ cells and tumor tissue may be used as a potential immunological prognostic indicator for the hepatocellular carcinoma patients after radical resection.
Zhonghua zhong liu za zhi [Chinese journal of oncology] 08/2008; 30(7):523-7.