Expression And Clinical Significance Of MiR-96and TACSTD1with Early Recurrence After Radical Surgery In Primary Hepatocellular Carcinoma

Background:Liver cancer is the one of most common malignancy worldwide currently in the global, cancer morbidity and mortality rates are among the forefront. China’s Ministry of Health statistics derived liver cancer mortality attributable to cancer mortality, especially, in second place. Primary hepatocellular carcinoma of liver cancer (hepatocellular carcinoma HCC), cholangiocarcinoma (cholangio carcinoma ICC) and mixed cell carcinoma. Hepatocellular carcinoma which accounted for more than90%. Surgery which is the traditional method of treating liver cancer is the preferred, but not all are suitable for patients with hepatocellular carcinoma, it is appropriate to treat with surgery Only cardiopulmonary function better and without metastasis. Most of patients with hepatitis, cirrhosis of the history, about80%of clinical patients for various reasons are inoperable. Non-surgical treatment of liver cancer varied, and each has its own method of treatment indications, only for the patient is the best way to approach. Should be based on the patient’s physical condition, liver function, of a tumor to select appropriate treatment.HCC recurrence is also possible, even underwent radical resection,which is related to the biological characteristics of HCC. Previous clinical studies have confirmed:the recurrence of hepatocellular carcinoma after curative resection is highest rate during the first year, and the prognosis of patients with relapse compared with patients with late recurrence is also poor. Therefore, early recurrence of HCC has become an important factor to affect the prognosis of patients. We must Find the key signaling pathways and target genes of early tumor recurrence to take effective measures to improve resectable liver cancer radical surgery rates and efficacy,which brings new hope for the treatment of clinical liver cancer.Our previous studies established a genomics research in line with specimens of liver cancer data repository and use AffymetrixU133plus2.0chips in genome-wide comparison of10cases of recurrence after one year and10cases more than two years without differences in gene expression profiles of liver cancer recurrence in two patients, found a number of sensitive genes associated with early recurrence of HCC. We have found that the significance of the four probes on the chip (probeset) both point discoidin domain receptor tyrosine kinase (discoidin domain receptor tyrosine kinase1, DDR1) gene; early recurrence group and the4probes were significantly up-regulated genes were significantly upregulated tips DDR1closely associated with early recurrence of HCC. Further RT-PCR and western-blot experiments also confirmed DDR1gene in patients with early recurrence of HCC was significantly higher in patients with non-early relapse, DDR1gene expression is closely related to the highly invasive carcinoma. This is highly suggestive, DDR1gene may become a biological gene therapy of liver cancer, a new sensitive molecular target.The previous researchers took advantage of miRNA chip (Agilent) technology compared recurrence after1year and not more than two years after the two groups of patients with hepatocellular carcinoma recurrence in genome-wide differences in miRNA expression profiling, discovery and early recurrence of HCC related to some MicroRNA differential expression of miR-96in which bioinformatics technology (Targetscan, RNA22, miRanda) may be associated with DDR1predict gene expression and miRNA microarray screening early recurrence of hepatocellular carcinoma is closely related miRNA, published " use chip technology liver cancer screening closely related to early recurrence microRNA ", studies have found a number of early recurrence group than in the non-regulated and down-regulated significant decrease in miRNA expression.Previous studies,miR-96in pancreatic cancer as a tumor suppressor gene KRAS miRNA targeted directly in clinical specimens, the measured expression of miR-96or disappear, and KRAS was upregulated by in vitro studies have shown that miR96KRAS down relevant way to reduce cancer cell invasion and migration, other studies have also found that miR-96in breast cancer, prostate cancer, since the negative regulatory role in the development.Preliminary studies using gene chip technology in the genome-wide detection of early recurrence were compared and early recurrence of HCC in patients with non-tumor tissue gene expression profiling, screening and early cancer recurrence sensitive genes related to, and published " using gene chip technology early liver cancer screening relapse-related genes ", the study found a number of early recurrence group than in the non-regulated and down-regulated genes expressed early recurrence group, these differentially expressed genes are mainly sports-related cell genes, signal transduction genes, cell cycle-related apoptosis gene and the like. Where in the tumor-associated calcium signal transducer1(TACSTD1) wherein the cell adhesion genes are sports-related gene is a cancer-associated antigen is expressed in endothelial cells and tumors of the gastrointestinal tract associated with cell adhesion, positioned at chromosome chr2p21, recent studies have found it there and induces the expression of cell proliferation by up-regulating c-myc, can be used as immunotherapy target molecules in other tumors such as esophageal cancer, cervical cancer were confirmed TACSTD1highly expressed in cancer tissue which, in this study, ER group compared with hER group TACSTD1gene expression was significantly increased, reaching more than eight times, suggesting that it is closely related to early recurrence of HCC, may be used as an early recurrence of HCC sensitive targets.Objective:Based on a large sample, we use real-time PCR technology and immunohistochemistry to find differences in expression levels of miR-96and TACSTD1between early postoperative recurrence group and non-group early recurrence in hepatocellular carcinoma, to explore the relationship between the expression of miR-96and TACSTD1early recurrence of hepato cellular carcinoma after curative.Methods:Based on screening, early recurrence is defined as the time from liver cancer recurrence after radical resection to tumor recurrence within one year, the definition of non-early HCC recurrence after radical more than two years and no recurrence of liver cancer.61cases were selected from Guangdong Provincial People’s Hospital studied, specimens are cryopreservation of tissue. Which early recurrence of HCC group of33patients (radical postoperative intrahepatic recurrence of lesions within1year),28cases of early non-recurrence group (after over two years of intrahepatic recurrence of lesions does not appear). We collected61cases of clinical data and tumor data including gender, age, tumor size,vascular invasion or not, intrahepatic metastasis or not, portal vein thrombosis or not, tumor grade and so on. Real-time PCR were used to verify the miR-96and immunohistochemical techniques to verify TACSTD1expression levels in the two HCC tissue.Using SPSS13.0software for statistical analysis, measurement data(x±s) said, using t test, count data using Fish’er exact test, X2t est, P<0.05was considered statistically significant.Results:1. Early recurrence after radical resection of hepatocellular carcinoma group in tumor diameter (P<0.001) were statistically significant; tumor size in this study may be a factor that affect the early recurrence after radical resection of hepatocellular carcinoma, while others such as age, gender, vascular thrombosis, intrahepatic metastasis, histological grade level, portal thrombosis and other factors need more further studies to conform.2. Real-time PCR results of miR-96in61cases of hepatocellular carcinoma showed early recurrence group relative to non-HCC[(0.634±0.783) VS(5.182±11.321), P=0.016], miR-96expression was significantly lower in the group of early recurrence in hepatocellular carcinoma, down8.174times of the two groups the difference is statistically significant.3. According to the median miR-96in HCC tissues relative expression values for the sector, the group was divided into61cases of liver cancer patients with high expression group and the low expression group, using statistical analysis, miR-96expression levels in HCC and the size of the tumor diameter (X2=4.761, P=0.029), early HCC recurrence (X2=4.867, P=0.027) related, there are significant differences, no significant difference in the rest of the features.4. TACSTD1in the positive control (esophageal carcinoma) was localized to the cell membrane, brownish yellow staining in normal liver tissue was negative, positive staining in this experiment located in the cell membrane;61cases of liver cancer tissues positive rate of8.2%(5/61), in which the expression was9.09%(3/33), the non-recurrence of early positive expression group was7.14%(2/28) in the early recurrence group; there are no significant differences in two groups (P=0.999).Conclusions:1. tumor size is a factor closely related to the early postoperative recurrence of liver cancer,while others such as age, gender, vascular thrombosis, intrahepatic metastasis, histological grade level, portal vein thrombosis and other factors need more further studies to conform;2. the expression of miR-96in the early recurrence of HCC is much lower than in the non-early recurrence group, which is closely related to the early recurrence after liver cancer, and miR-96expression in HCC is related to the the diameter of the tumor, liver surgery early recurrence, miR-96may predict cancer recurrence after radical resection of liver cancer;3. We can not determined the relationship between TACSTD1and early recurrence after radical and look forwad to further studies.