Huanping Lin

Correlation between Clinicopathology and Expression of Heat Shock Protein 72 and Glycoprotein 96 in Human Esophageal Squamous Cell Carcinoma

Heat shock protein 72 (HSP72) and glycoprotein 96 (gp96) are highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 and gp96 in the development and progression of gastrointestinal carcinomas but detailed information is still ambiguous. We investigated the correlation between clinicopathology and expression of HSP72 and gp96 in human esophageal squamous cell carcinoma. The expression of HSP72 and gp96 was studied in 120 human esophageal squamous cell carcinomas with or without metastasis as well as in mucous membrane adjacent to cancers by way of immunohistochemistry. HSP72 immunoreactivities were detected in 112 of 120 primary tumors (93.3%) and in 30 of 120 mucous membranes adjacent to cancers (25.0%). Gp96 detected in esophageal squamous cell carcinoma and inmucous membrane adjacent to cancer was 85.0% and 20.0%, respectively. Both HSP72 and gp96 stained in cytoplasm. HSP72 and gp96 expression in esophageal squamous cell carcinomas withmetastasis was significantly higher than those with nonmetastasis (P < .05). The results indicate that there exists a significant correlation between the expression of HSP72 and gp96 and the progression of esophageal squamous cell carcinomas. HSP72 and gp96 expression were significantly associated with the presence of tumor infiltration, lymph node, and remote metastasis.

Recombinant heat shock protein 70 functional peptide and alpha-fetoprotein epitope peptide vaccine elicits specific anti-tumor immunity

Alpha-fetoprotein (AFP) is a marker of hepatocellular carcinoma (HCC) and serves as a target for immunotherapy. However, current treatments targeting AFP are not reproducible and do not provide complete protection against cancer. This issue may be solved by developing novel therapeutic vaccines with enhanced immunogenicity that could effectively target AFP-expressing tumors. In this study, we report construction of a therapeutic peptide vaccine by linking heat shock protein 70 (HSP70) functional peptide to the AFP epitope to obtain HSP70-P/AFP-P. This novel peptide was administered into BALB/c mice to observe the effects. Quantification of AFP-specific CD8 + T cells that secrete IFN-γ in these mice via ELISPOT revealed the synergistic effects of HSP70-P/AFP-P with increased numbers of AFP-specific CD8 + T cells. Similarly, ELISA analysis showed increased granzyme B and perforin released by natural killer cells. Moreover, in vitro cytotoxic T-lymphocyte assays and in vivo tumor preventive experiments clearly showed the higher antitumor effects of HSP70-P/AFP-P against AFP-expressing tumors. These results show that treatment of BALB/c mice with HSP70-P/AFP-P induced stronger T-cells responses and improved protective immunity. Our data suggest that HSP70-P/AFP-P may be used as a therapeutic approach in the treatment of AFP-expressing cancers.

HSP72 and gp96 in gastroenterological cancers

Heat shock protein 72 (HSP72) and glycoprotein 96 (gp96) are highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 and gp96 in the development and progression of gastrointestinal carcinomas but detailed mechanisms are still ambiguous. Human esophageal cancer, gastric cancer, colon cancer and liver cancer are common gastrointestinal malignant carcinomas in the world. The studies indicated that there existed a significant correlation between the expression of HSP72, gp96 and the development and progression of digestive carcinomas. HSP72 and gp96 expression were significantly associated with the presence of tumor infiltration, lymph node and remote metastasis. Interestingly, studies have found that HSP72 chaperoned alpha-fetoprotein (AFP), HBx in hepatocellular carcinoma, and CD44 in colonic carcinomas. The further researches demonstrated that HSP72-AFP or gp96-AFP recombined vaccine could elicit specific anti-tumor immunity. The high-level expression of HSP72 and gp96 may be not only used as diagnostic or prognostic markers for gastrointestinal carcinomas but also as better immunotherapeutic vaccines in the cancers.

Specific Antitumor Immunity Induced by Cross-linking Complex Heat Shock Protein 72 and Alpha-fetoprotein

Hepatocellular carcinoma (HCC) is one of the most common malignancies over the world. Alpha-fetoprotein (AFP) is an oncofetal protein during HCC development that could generate weaker and less reproducible antitumor protection, and it may serve as a target for immunotherapy. Therefore, it is imperative to enhance its immunogenicity and develop therapeutic vaccines to eliminate AFP-expressing tumors. In this study, by way of glutaraldehyde cross-linking, we constructed a potential therapeutic protein complex vaccine, heat shock protein 72 (HSP72)/AFP. Our results demonstrated that AFP and HSP72 synergistically exhibited significant increases in AFP-specific CD8(+) T cell responses and impressive antitumor effects against AFP-expressing tumors. Priming mice with the reconstructed vaccine, we elicited robust strong protective immunity. Our study suggests that a tumor vaccine by cross-linking tumor antigen and HSP72 is a promising approach for cancer therapy.

Glycoprotein 96 and α-Fetoprotein Cross-Linking Complexes Elicited Specific Antitumor Immunity

Hepatocellular carcinoma (HCC) is one of the most common malignant gastroenterological cancers over the world. α-fetoprotein (AFP) is an oncofetal protein produced during HCC development that could generate weaker and less reproducible antitumor protection, and it may serve as a target for immunotherapy. Therefore, it is imperative to enhance its immunogenicity and develop therapeutic vaccines to eliminate AFP-expressing tumors. In this study, by way of glutaraldehyde cross-linking, we constructed a potential therapeutic protein vaccine, glycoprotein 96 (gp96)/AFP. Our results demonstrated that AFP and gp96 synergistically exhibited significant increase in AFP-specific CD8⁺ T-cell responses and impressive cytotoxic antitumor effect against AFP-expressing tumors. Priming mice with the reconstructed vaccine, we elicited robust strong protective immunity. Our study suggests that tumor vaccine by cross-linking tumor antigen and gp96 is a promising approach to cancer therapy.

Complex Formation between Heat Shock Protein 72 and Hepatitis B Virus X Protein in Hepatocellular Carcinoma Tissues

Hepatitis B virus (HBV) infection is a major factor contributing to the development of hepatocellular carcinoma (HCC) in the world. Hepatitis B virus X protein (HBx) has been verified to play an important role in hepatocarcinogenesis. Heat shock protein 72 (HSP72) as a molecular chaperone has been confirmed to overexpress in epithelial carcinoma cells. There may be a possible association between the expression of HSP72 and HBx during the growth and progression of hepatocellular carcinoma cells. The aim of the study was to investigate the relationship between heat shock protein 72 and HBx in human hepatocellular carcinomas. The localization of HSP72 and HBx in human hepatocellular carcinomas was determined by immunohistochemistry and confocal laser microscopy. The interaction between HSP72 and HBx in liver cancer cells was analyzed by immunoprecipitation and Western blot. Our results revealed that hepatocellular carcinoma synchronously co-expressed higher level of HSP72 and HBx than that in the adjacent tissues to hepatocellular carcinoma. HSP72 and HBx were mainly immunolocalized in the cytoplasm. On the basis of immunoprecipitation and Western blot, we found that HSP72 formed complex with HBx in human hepatocellular carcinoma cells. The association between HSP72 and HBx in human hepatocellular carcinoma cells may contribute to study the pathogenesis and immunotherapy of hepatocellular carcinoma.

Heat shock protein 72 associated with CD44v6 in human colonic adenocarcinoma.

CD44v6 is a splice variant of CD44 (CD44v), probably promoting cancer cell adherence to vascular endothelium and base membranes and enhancing the invasion and metastasis of colonic carcinomas. Heat shock protein 72 (HSP72) as a molecular chaperone has been confirmed to be overexpressed in epithelial carcinoma cells. There may be a possible association between the expression of HSP72 and CD44v6 during the growth and progression of colonic carcinoma cells. The aim of the study was to investigate the interaction between heat shock protein 72 and CD44v6 in human colonic carcinomas. The localization of HSP72 and CD44v6 in human colonic carcinomas was determined by immunohistochemistry and confocal laser microscopy. The interaction between HSP72 and CD44v6 in colonic carcinoma cells was analyzed by immunoprecipitation and Western immunoblots. Our results revealed that colonic carcinoma synchronously co‐expressed higher levels of HSP72 and CD44v6 than that in adjacent normal colonic tissues. HSP72 and CD44v6 were mainly immunolocalized in the cytoplasm, and also immunolabelled on the cell membrane. Based on immunoprecipitation and Western immunoblots, we found that HSP72 was associated with CD44v6 precursor fragments in human colonic carcinoma cells. The interaction between HSP72 and CD44v6 in human colonic carcinoma cells may contribute to study the pathogenesis and immunotherapy of colonic carcinoma.

Mechanisms of Biotherapy Effect of Shenfoweikang Herbs on Gastric Carcinoma Cells

To explore the biotherapy effect of Shenfoweikang herbs in treatment of gastric cancer, BALB/C mice were grafted with a mouse gastric adenocarcinoma cell line MFC as the experimental model. The mice were divided into four groups. Mice in the experimental groups received different doses of Shenfoweikang herbs over a 60-day period starting at the first day after grafting. Mice received saline as controls. All the mice were sacrificed at 61 days after being grafted. Tumor size was periodically measured during the life of the mice and tumor weight was determined by an electron balance immediately after the animals killed. Serum cytokines, granzyme B and perforin were examined by the ELISA method. The anti-tumor effect was detected by the cytotoxic T-lymphocyte (CTL) assays. Our results demonstrated that Shenfoweikang herbs could inhibit the growth of gastric cancer by activating the CTLs and inducing the secretion of cytokines, perforin and granzyme B. Our study suggests that Shenfoweikang herbs inhibited the proliferation of gastric carcinoma by activating the effects of immune cells, which may lay a better basis for further study on gastric cancer biotherapy.

Bioinformatics of Rhizoma Curcumae against Gastric Carcinoma Grafted on Mice

Objective: The study is intended to explore the anti-tumor effect of the rhizoma curcumae in treatment of gastric cancer. Methods: Kunming mice grafted with a mouse gastric adenocarcinoma cell line MFC was used as the experimental model. The mice were divided into four groups, one control and the other three different drug administration experimental groups. The anti-tumor effect was assessed by tumor size, apoptotic indices (AI) and the expression of VEGF, STAT3 and HIF-1α in tumor tissues. Results: Compared with controls, tumor size and weigh were significantly inhibited with the rhizoma curcumae decoction treatment (P<0.05). AI in gastric cancer grafted mice was significantly increased in the rhizoma curcumae decoction group. The expression of VEGF, STAT3 and HIF-1α in tumor tissues were down-regulated after treated with rhizoma curcumae decoction. Conclusion: The anti-tumor effect of gastric cancer cell growth in vivo by rhizoma curcumae is related with the induction of the cell apoptosis and down-regulation of VEGF, STAT3 and HIF-1α signal transduction system.

Chaperoned function of HSP72 in hepatocellular carcinomas

Heat shock protein 72 (HSP72) is highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 in the development and progression of gastrointestinal carcinomas but detailed mechanisms are still ambiguous. Human hepatocellular carcinomas are common gastrointestinal malignant carcinomas in the world. The studies indicated that HSP72 chaperoned alpha-fetoprotein (AFP) and HBx in hepatocellular carcinoma. The further researches demonstrated that HSP72-AFP recombined vaccine could elicit specific anti-tumor immunity. The high-level expression of HSP72 may be not only used as diagnostic or prognostic markers for hepatocellular carcinomas but also as better immunotherapeutic vaccines in the cancers.