Expressions Of Snail,VEGF,MMP-9mRNA And Protein In Hepatocellular Carcinoma

Objective To investigate the hepatocellular carcinoma tissues Snail, VEGF, MMP-9 expression of three biological indicators and analyze the three biological indicators of the correlation between expression and clinical pathological correlation and its hepatocellular carcinoma invasion and metastasis of the role, and its expression in cancer invasion and metastasis of liver cancer for the prediction, the degree of malignancy to determine whether the role of important tips.Methods In this paper, VP-9000 two-step immunohistochemistry to detect 70 cases of HCC and its adjacent tissues in the Snail, VEGF, MMP-9 protein expression, using RT-PCR, detected 40 cases of fresh tumor tissue and its adjacent tissues Snail, VEGF, MMP-9mRNA expression,30 cases of carcinomas were selected from the 2007-2009 hospital after surgical resection and pathological diagnosis of primary hepatocellular carcinoma patients, paraffin blocks per case contain cancer and adjacent tissues.22 patients were male and 8 female. Aged 36-77 years (median age 56.5 years). HCC Edmondson grade 1 cases of gradeⅠ,Ⅱgrade 4 cases,Ⅲgrade 20 cases,Ⅳlevel in 5 cases.40 cases of fresh tumor tissue removed at surgery from the edge of 3～5 cm tumor adjacent tissue removed, and then take no hemorrhage, necrosis, and actively growing tissues, respectively-70℃refrigerator for preservation, for the RT-PCR, the remaining organization with 40 g/L paraformaldehyde, paraffin embedded sections for immunohistochemistry.Results immunohistochemistry staining intensity according to the depth of integration (P),0 points:no staining,1 point:weak positive,2 points:strongly positive. By color cell percentage (I), 1 point:0-25%,2 points:26%-50%,3 points 51%-75%,,4 points: 76%-100%, and then follow the positive cells×staining intensity, calculate the total score, total score≤3 were negative,≥3 were positive. Snail mainly expressed in the nucleus, Snail positive expression rates in HCC was higher than adjacent tissues (P<0.05), and Snail expression in the cancer rate have relationship with capsule invasion, portal vein invasion and intrahepatic metastasis (P <0.05),VEGF mainly in the cytoplasm, HCC positive expression of VEGF was higher than in adjacent tissues (P<0.05), and the cancer rate of VEGF expression have relationship with tumor diameter, portal vein invasion, capsule invasion and intrahepatic metastasis (P <0.05). MMP-9 mainly in the cytoplasm, positive expression of MMP-9 was higher in HCC than in adjacent tissues (P<0.05), and the rate of MMP-9 expression in cancer have relationship with tumor size, portal invasion, capsule invasion and intrahepatic metastasis (P<0.05). Snail, VEGF and MMP-9 expression in hepatocellular carcinoma by using x2 test, Snail and VEGF Was a positive correlation, (P<0.05); Snail and MMP-9 was a positive correlation, (P<0.05). RT-PCR product with 1.5% agarose gel electrophoresis and observe the photo, PCR products corresponding specific bands, as the positive results of hepatocellular carcinoma。SnailmRNA positive expression rates was higher than adjacent tissues (P<0.05), and cancer SnailmRNA positive expression rate have relationship with capsule invasion, portal invasion case the (P <0.05), but has nothing to do with intrahepatic metastasis (P<0.05) positive hepatocellular carcinoma VEGFmRNA expression rates was higher than cancer adjacent tissues(P<0.05), and cancer SnailmRNA positive expression rate have relationship with capsule invasion, portal invasion, intrahepatic metastasis related (P<0.05). HCC MMP-9mRNA expression higher than that in adjacent tissues (P< 0.05), and cancer MMP-9mRNA positive expression rate have relationship with capsule invasion, portal invasion and intrahepatic metastasis (P<0.05) Snail, VEGF and MMP-9mRNA in hepatocellular carcinoma by using x2 test, SnailmRNA and VEGFmRNA expression were positively correlated, (P<0.05); Snail mRNA and MMP-9mRNA positive correlation, (P<0.05).Conclusion of the present study, immunohistochemistry and PCR detection of hepatocellular carcinoma and adjacent tissues Snail, VEGF, MMP-9 mRNA and protein expression, obtained Snail, VEGF, MMP-9mRNA and protein were highly expressed in hepatocellular carcinoma, low expression in cancer adjacent tissues. Snail, VEGF, MMP-9 protein expression in HCC have relationship with the capsule invasion, portal vein invasion and intrahepatic metastasis, but has nothing to do with tumor size, tumor grade and tumor, generally showing that Snail, VEGF, MMP-9 in the liver cancer invasion and metastasis play a role, Snail, VEGF, MMP-9 protein expression enhances the cancer cell line with high invasive Make it more susceptible to invasion and metastasis. Snail at the mRNA level and the capsule invasion, portal invasion but did not show correlation with intrahepatic metastasis, suggesting that sample size may be less relevant, it may raise levels of Snail protein in translation or increased stability of the protein. Only the inhibition of E-cad expression were not enough Snail in tumor invasion and metastasis, the Snail pathway are also involved in other protein molecules, some studies have shown that VEGF abroad is in the upper reaches of the gene product, induced Snail gene promoter expression, the expression of Snail can further control after its downstream genes, such as E-cadherin, MMP expression, Snail expression can be regulated MMPs, MMPs can degrade extracellular matrix to promote tumor metastasis. This requires more research to fully understanding the role of Snail in HCCs and different target genes. Studies have demonstrated antisense Snail transcription factor can lead to liver cancer cells in vitro invasion and motility potential decline, suggesting that Snail signaling cascade living in the central link block Snail signaling pathway, inhibit the expression of Snail, is expected to become the treatment of liver cancer new target.