Expression And Clinical Significance Of Tcell Immunoglobulin3(TIM-3) In Hepatocellular Carcinoma

HCC (hepatocellular carcinoma, HCC) is one of the most common malignant tumor in the world. In some developed countries, hepatocellular carcinoma is the main tumor type and has high incidence and cancer mortality around the world. As a whole, in the most common malignant tumor, men’s incidence ranks seven, while women’s ranks nine. In our country, the annual death rate of liver cancer accounts for the second in cancer mortality. In the past20years, increasing incidence and younger and diversification trend of HCC make people feel worried. Those patients without nonspecific symptoms at early stage had mostly lost the opportunity of surgical therapy when liver cancer was detected at advanced stage. Besides, the biological malignant degree of HCC is high,5-year survival rate is low, and the prognosis is poor overall. In recent years, imaging diagnosis techniques and molecular diagnosis technology have greatly increased the rate of early detection of HCC. With the development of diagnostic imaging and molecular diagnostic techniques, the rate of early diagnosis of primary liver cancer has significantly improved. The previous treatment simply by surgery alone has been multiplied into comprehensive treatment such asradio-frequency ablation, transcatherter arterial chemoembolization (TACE), radiotherapy, chemotherapy, Chinese medicine and medical therapy, immunotherapy, molecular targeted therapy and gene therapy, even liver transplantation but the curative effect are not satisfactory. Its pathogenesis and related treatment also need further in-depth study. Studies have shown that hepatocellular carcinoma patients have obvious cellular immune dysfunction and lower anti-tumor immune function. It is alleged that cellular immunity is the core of the antitumor immunity and humoral immunity is only the synergy in some cases. Due to the important role of cellular immunity in tumorigenesis, the relationship between cellular immune and hepatocellular carcinoma (HCC) becomes the focus of basic and clinical research of liver cancer. It is generally acknowledged that T cellular immunity is the core of the antitumor immunityand CD8+T cells can directly specificity kill cancer cells, known as killer T cells (CTL). CD4+T cells also play an important role in regulating tumor immunity. For advanced cancer patients whose Thl cells is restrained, if the Thl/Th2balance state tends to Thl deviation, the tumor immunity may be adjusted.T cell immunoglobulin and mucin-domain containing molecule (TIM) gene family is discovered in2001when Mclntire searched for asthma susceptibility genes in the mice and is identified to be new gene family after the analysis of the genome and cloning location. The gene family structure contains immunoglobulin V and mucins area, the protein of TIM gene encoded is type I transmembrane glycoprotein with a common motif, and the basic structure consists of a signal peptide, immunoglobulin, sticky protein sample region, transmembrane region and phosphorylation sites in the intracellular region. TIM gene is located on mice chromosome llbl.land consists of eight members in mice, encoding TIM-1-TIM-8. Human TIM gene families have a high homology with mice which is located on human chromosome5q33.2and consists of three members in human, encoding TIM-1, TIM-3and TIM-4.TIM-3is the important member of TIM family, It was originally identified as a mouse Thl-specific cell surface marker that was expressed after several rounds of Thl differentiation in vitro. TIM-3is expressed on a subset of CD4+T cells, on CD8+T cells, on Thl7cells Treg cell, natural killer cells, dendritic cells and monocytes/macrophages, with the deepening of the study, the gene structure, biological characteristics and role in the immune regulation function are more and more clear.At present, the study of TIM-3in tumor immune mainly concentrates in patients with snuff cancer, breast cancer, melanoma skin cancer, hepatocellular carcinoma and so on, but the research with TIM-3in HCC patients is rarely reported. As the HCC has highly malignant biological characteristics and is different from the above several kinds of tumors, the research the expression of TIM-3in hepatocellular carcinoma is necessary in order to further explore whether TIM-3is involved in the formation and development of HCC. This study is to detect the expression of TIM-3in tumor tissues, para-carcinoma tissue, normal liver tissues and peripheral blood mononclear cells of HCC patients with flow cytometry and routine PCR and explore the relationship between HCC and the above.Objectives:Detect the expressions of TIM-3mRNA in tumor tissues of patients with HCC by RT-PCR and detect the expressions of TIM-3protein in CD4+Tcell, CD8+T cell of tumor tissues and peripheral blood, explore the relationship between HCC and TIM-3.Methods:Detect the expression of TIM-3and TIM-3mRNA from16human liver specimens in tumor tissues, para-carcinoma tissues and peripheral blood mononclear cells of HCC patients with flow cytometry and routine PCR. Detect the expressions of TIM-3and TIM-3mRNA in5cases of healthy volunteers’peripheral blood,5cases of patients with gallbladder stones and5cases of liver hemangioma patients’normal liver tissue, using as a control group.Results1The results of RT-PCR technology showed that the TIM-3mRNA expressed in liver cancer patients group is obvious and expressed in para-carcinoma tissue is weak. The expression of TIM-3in normal liver tissues was undetectable.2The level of TIM-3expression on healthy volunteers and HCC patients’peripheral blood is no express. In hepatocellular carcinoma patients’peripheral blood, CD4+Tcell is expressed as (0.20%±0.04%) and CD8+T cell is expressed as (0.19%±0.03%). In healthy volunteers’ peripheral blood,CD4+T cells is expressed as(0.08%±0.02%)and CD8+T cells is expressed as(0.11%±0.08%).In short,there was no statistically significant difference.3The level of TIM-3expression on gallbladder stones and liver hemangioma patients’normal liver tissue is also no express.In normal liver tissue,CD4+T cells is expressed as (0.18%±0.04%)and CD8+T cells expressed as(0.21%±0.03%).The level of TIM-3expression is higher in hepatocellular carcinoma tissues.TIM一3in cancer CD4+T cells expressed as(3.53%±0.54%)and CD8+T cells is expressed as(3.36%±0.27%)；for TIM一3in para-carcinoma tissue,CD4+T cells is expressed as (1.73%±0.16%),and CD8+T cells is expressed as1.82%±0.37％).The level of TIM-3expression in cancer tissues of CD4+T cell and CD8+T lymphocytes was significantly higher than that in para-carcinoma tissue and normal liver tissue,and all have significant difference(P＜0.01).Conclusions:1The TIM-3mRNA expressed in tumor tissues of patients with HCC is obvious increased.2The TIM-3is highly upregulated on both CD4+T cell and CD8+Tcell from HCC cancer tissues.3In the occurrence and development of hepatocellular carcinoma (HCC),TIM-3May take part in antitumor immunity by regulating T cell immune functions, which could provide new targets for clinical immunotherapy.