Though considerable efforts aim at elucidating the tumorigenesis of ovarian carcinoma, its molecular mechanism has not been completely explained. Recently, MACC1 has been identified as a prognosis biomarker for colon cancer, which promotes proliferation, invasion and hepatocyte growth
factor (HGF)-induced scattering of colon cancer cells in vitro and in vivo [2]. RG7112 manufacturer MET, which encodes Met protein, has been proven to be a transcriptional target of MACC1. MACC1 controls the activity and expression of MET, and regulates HGF/Met signal pathway [2]. HGF/Met pathway plays key roles in carcinogenesis, aberrant activation of Met leads to enhancement of cell proliferation, invasion and metastasis, and Met is essential for metastatic potential of many malignances [3]. Once activated by HGF, Met transmits Y-27632 solubility dmso intracellular signals and activates downstream Ras-mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/Akt pathways, which promote cell survival, migration, invasion, and suppress apoptosis [4]. MACC1 was demonstrated to be associated with poor prognosis and high risk of metastasis in colon cancer, gastric carcinoma, lung cancer, and hepatocellular carcinoma [5–8].
However, the mechanism of MACC1 implicates in ovarian cancer is still unclear. Small interfering RNA can specifically silence particular genes, and is used as a powerful tool to research gene functions and as a genetic therapy strategy for carcinoma [9]. In present study, expressions of MACC1 were detected in different ovarian tissues by immunohistochemistry, effects
of MACC1 inhibition on OVCAR-3 cells were observed by RNA interference, and the possible antitumor mechanisms of MACC1 knockdown in ovarian carcinoma cells were discussed. Materials and methods Immunohistochemistry and evaluation Paraffin-embedded 20 specimens of normal ovary, 19 specimens of benign ovarian tumor and 52 specimens of ovarian cancer tissues were obtained from Department of Pathology of Zhengzhou University. Rabbit-anti-human polyclonal MACC1 antibody (Sigma, USA) was used for immunohistochemistry assay, which was performed following the protocol of Universal SP kit (Zhongshan Goldenbridge Biotechnology, Peking, China). Positive staining of MACC1 protein presents Aspartate brown in cytoplasm, partly in nucleus. Semi-quantitative counting method was used to determine positive staining described as following: this website Selected 10 visual fields under high power lens (× 400) randomly, counted the numbers of positive cells in 100 cells per field, calculated the average positive rate. Positive rate less than 1/3 scored as 1, more than 1/3 and less than 2/3 scored as 2, more than 2/3 scored as 3, without positive cell scored as 0. Cells without brown staining scored as 0, with mild brown staining scored as 1, with moderate brown staining scored as 2, with intense brown staining scored as 3.
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