The Expression And Significance Of The ANXA-Ⅱ,VEGF In HepG2 Cells Under The Disturbance Of 5-fluorouracil

AIMPrimary carcinoma of liver is one of the malignant tumor with high fatality rate, the death toll of liver cancer of our country is the most of the world,90% of which is hepatocellular carcinoma (HCC), the key of the poor treatment effect and high fatality rate is the high power of attack and metastasis of hepatoma carcinoma cell. At present, the molecular mechanism of attack and metastasis of tumor cell is one of the focuses in the study of tumor therapeutics. The generation of renascent vessels of tumor, the degradation of the base material out of the cells and adhesion of the cells are the main mechanisms which are investigated deeply presently. It has been testified that ANXA and Vascular endothelial growth factor (VEGF) play important roles in the mechanism of attack and metastasis of tumor, but the specific mechanism and the interaction between them in it have not been known. ANXA2 is an important member in ANXA, and it exerts important function in the mechanism of attack and metastasis of many tumors, such as colorectal cancer, lung cancer, stomach cancer, esophageal cancer, carcinoma of prostate, leukemia, multiple myeloma, cervical carcinoma, thyroid carcinoma. For the moment, the research of the expression of ANXA2 in liver cancer is very few. In vivo, VEGF is the most powerful vascular growth factor, which participates in and induces generation of renascent vessels of tumor directly. It has been approved that VEGF and its receptor VEGFR express much more strongly in the tissue of liver cancer than that in the tissue of normal liver, they exert important function in the growth, metastasis, recrudescence of liver cancer. In this research, under disturbance of different dose of 5-fluorouracil (5-FU), we discuss the pertinence and significance of the expression of ANXA2,VEGF, by detect the expression of them in HepG2 liver cancer cells and the power of attack and metastasis and the change in energy of growth of the HepG2 liver cancer cells,which provides a new way for illuminating the molecular mechanism of attack and metastasis in HCC, the direction of therapy and the judgement of prognosis.MethodsBy MTT, Transwell, Westernblot, RT-PCR, immunohistochemistry, under disturbance of different dose of 5-fluorouracil (1:blank group,2:50 ug/ml,3:100 ug /ml,4:200ug/ml,5:400ug/ml), detect the expression of ANXA-2,VEGF in HepG2 liver cancer cells and the pertinence between them, at the same time, observe the change in power of attack and metastasis and energy of growth of the HepG2 liver cancer cellsResultsIn MTT, the rate of restraint of the HepG2 liver cancer cells growth increase gradually as the dose of 5-fluorouracil increase, the discrepancy between the groups of different dose have significance of statistics(P<0.05);In Transwell, the invasiveness of HepG2 liver cancer cells fall evidently as the dose of 5-fluorouracil increase, the discrepancy between the groups of different dose have significance of statistics(P<0.05);In RT-PCR, the expression of the mRNA of ANXA2,VEGF in the HepG2 liver cancer cells decrease gradually as the dose of 5-fluorouracil increase, the discrepancy between the groups of different dose have significance of statistics (P<0.05), and the expression between them have notable positive pertinence, the coefficient of spearman correlation is rp=0.900;In Westernblot, the expression of the protein of ANXA2,VEGF in the HepG2 liver cancer cells decrease gradually as the dose of 5-fluorouracil increase, the discrepancy between the groups of different dose have prominent significance of statistics(P<0.01), and the expression between them have notable positive pertinence, the coefficient of spearman correlation is rp=0.856; In immunohistochemistry, the expression of ANXA2,VEGF in the HepG2 liver cancer cells decrease gradually as the dose of 5-fluorouracil increase, the discrepancy between the groups of different dose have significance of statistics (P<0.05).ConclusionAs the dose of 5-fluorouracil increase, the power of attack and energy of growth of the HepG2 liver cancer cells decrease gradually, the expression of mRNA and protein of ANXA2,VEGF decrease gradually, the expression between them have positive pertinence, it implies that 5-FU restrain the attack and metastasis of the HepG2 liver cancer cells maybe by the way of ANXA2,VEGF, they might cooperate with each other to promote attack and metastasis in hepatocellular carcinoma.