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An Experimental Research About The Effect Of MiRNA-141 The On The Sensitivity Of Chemotherapies And The Progression Of The Colon Cancer By Targeting The MAP4K4

Posted on:2017-02-10Degree:MasterType:Thesis
Country:ChinaCandidate:F F WangFull Text:PDF
GTID:2334330485973945Subject:Surgery
Abstract/Summary:PDF Full Text Request
Objects:To investigate the effect of miRNA-141 on the proliferation, invasion and metastasis of colon cancer cells, to explore the possible molecular mechanism inhibiting the colon cancer cells, and to study the effect of the combination of miRNA-141 and chemotherapy drugs on the colon cancer cells.Methods:1 The expression levels of miRNA-141 in colon cancer cells being treated with miRNA-141 mimics and miRNA-141 inhibitor were detected by qPCR.2 The proliferative activity, invasion ability and migration ability of the colon cancer cells with miRNA-141 over-expression or low-expression were detected by MTS, Transwell Matrigel and wound scrape assays.3 The expression levels of MAP4K4 genes of the colon cancer cells with miRNA-141 over-expression or low-expression were detected by qPCR.4 The expression of MAP4K4 proteins of the colon cells with miRNA-141 over-expression or low-expression were detected by Western blotting assay.5 Real-time fluorescent quantitative PCR detected the expression of miRNA-141 in HCT-116 and HCT-8 colon cancer cells being treated with Oxaliplatin(25mg/l, 12.5mg/l, 0mg/l) and Fluorouracil(25mg/l, 12.5mg/l, 0mg/l), respectively.6 The expression levels of MAP4K4 genes in colon cancer cells after being treated with these two drugs were detected by RT-PCR.7 The effect of Fluorouracil and Oxaliplatin on colon cancer cells proliferation activity with different concentrations(25mg/l, 12.5mg/l, 0mg/l)were detected by MTS assay.8 The invasion ability and migration ability of the colon cancer cells after being transfected with miRNA-141 mimics, miRNA-141 mimics control,the two drugs and the combination of miRNA-141 mimics and drugs were detected by Transwell Matrigel and wound scrape assays, respectively.9 The expression levels of MAP4K4 genes of the colon cancer cells after being transfected with miRNA-141 mimics, miRNA-141 mimics control, the two drugs and the combination of miRNA-141 mimics and drugs were detected by RT-PCR, respectively.10 The expression levels of MAP4K4 proteins of colon cells after being treated with the two drugs, transfected with miRNA-141 mimics, transfected with miRNA-141 mimics control and treated with miRNA-141 mimics and drugs were detected by Western blotting assay, respectively.Results:1 qPCR results revealed that the expression levels of miRNA-141 were decreased in colon cells being treated with inhibitor miRNA-141(P<0.05) and were raised in colon cancer cells being treated with miRNA-141 mimics(P<0.05).2 MTS assay indicated proliferative activity of colon cancer cells were inhibited after being transfected with miRNA-141 mimics but were advanced after being transfected with miRNA-141 inhibitor(P<0.05).3 Transwell Matrigel and wound scrape assays revealed the invasion ability and migration ability of colon cancer cells were inhibited after being transfected with miRNA-141 mimics but were promoted after being treated with miRNA-141inhibitor(P<0.05).4 qPCR results announced the expression levels of MAP4K4 genes of colon cancer cells were high after being treated with the miRNA-141 inhibitor but were low after being treated with the miRNA-141 mimics(P<0.05).5 Western blotting assay indicated the expression of MAP4K4 proteins of colon cancer cells were depressed after being treated with miRNA-141 mimics(P<0.05) but were increased when being treated with miRNA-141 inhibitor.6 qPCR results showed the expression levels of miRNA-141 in colon cancer cells being treated with these two drugs were increased(P<0.05).7 RT-PCR results showed the levels of MAP4K4 in colon cancer cells after being treated with these two drugs was reduced(P<0.05).8 MTS assays indicated that Fluorouracil and Oxaliplatin could depress the proliferation ability(P<0.05) of colon cancer cells, which can change by time and doses.9 Transwell Matrigel and wound scrape assays revealed the invasion ability and migration ability of colon cancer cells after being treated with the miRNA-141 mimics and drugs were inhibited significantly(P<0.05)10 RT-PCR results showed compared with the two drugs groups, treated with miRNA-141 mimics group and treated with miRNA-141 mimics control group, the expression of MAP4K4 of colon cancer cells after being treated with the miRNA-141 mimics and drugs were inhibited significantly(P<0.05).11 Western blotting assay results indicated compared with the the two drugs group, being transfected with miRNA-141 mimics and being transfected with miRNA-141 mimics control, the expression levels of MAP4K4 proteins of colon cancer cells being treated with the miRNA-141 mimics and drugs were inhibited significantly(P<0.05).Conclusions:1 The proliferative ability, invasion ability and migration ability of colon cancer cells were decreased after being treated with miRNA-141 mimics, so the miRNA-141 could suppress the progression of colon cancer.2 The expression of MAP4K4 genes were decreased in the colon cancer cells after being treated with miRNA-141 mimics. MAP4K4 was functioning as a tumor promoter.3 The proliferative ability of colon cancer cells was decreased by the two drugs.4 The invasion ability and migration ability of colon cancer cells after being treated with miRNA-141 mimics and the two drugs were inhibited.5 MAP4K4 proteins and genes of colon cells after being treated with miRNA-141 mimics and these two drugs were decreased. So the effects of miRNA-141 mimics on colon cancer cells was the same as the two drugs.6 The miRNA-141/MAP4K4 pathway can regulate the sensitivity of colon cancer cells to chemotherapeutic drugs.
Keywords/Search Tags:Colon cancer cell, MiRNA-141, MAP4K4, Fluorouracil, Oxaliplatin, Chemosensitivity
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