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MIR-130b-5p/RASAL1 Axis Affects Proliferation Of Gastric Cancer Cells Via RAS Pathways

Posted on:2018-01-02Degree:MasterType:Thesis
Country:ChinaCandidate:Y Q YangFull Text:PDF
GTID:2334330542951883Subject:Digestive medicine
Abstract/Summary:PDF Full Text Request
Objective:To testify the targeted relationship between miR-130b-5p and RASAL1 gene and intensive study the function of miR-130b-5p in gastric cancer in cell proliferation,migration and invasion.Methods:Expression of miR-130b-5p and RASAL1 in seven gastric cell lines were detected by qRT-PCR(Quantitative reverse transcription-polymerase chain reaction).MGC803 cells were selected for further study on account of obviously increased expression of miR-130b-5p accompanied with significantly decreased expression of RASAL1.MGC803 cells were transfected with miR-130b-5p mimics and miR-130b-5p inhibitor via Lipofectamine2000 for over and lower expression,with cells transfected with NC as control.In addition,a luciferase reporter gene assay was applied to testify the targeted relationship between miR-130b-5p and RASAL1.Then,the alternation of RASAL1 expression was detected by qRT-PCR and western blot analysis after transfected with miR-130b-5p mimics and miR-130b-5p inhibitor.In the end,cell proliferation,colony formation,migration and invasion ability were detected by cell proliferation assay,colony formation assay,and transwell assays,respectively.Results:Expression of miR-130b-5p was obviously increased in gastric cell lines accompanied with decreased level of RASAL1.RASAL1 was proved to be a target gene of miR-130b-5p by using Luciferase reporter gene assay.In addition,the expression of RASAL1 was lower in MGC803 cells that transfected with miR-130b-5p mimics and higher expression was detected in cells transfected with miR-130b-5p inhibitor in comparison with cells transfected with NC(control group)(P<0.05).Beyond that,the experimental group transfected with miR-130b-5p mimics manifested higher cell proliferation,strong ability of colony formation,superior ability of migratory and invasion,and to the contrary,cells transfected with miR-130b-5p inhibitor show weaker ability in proliferation,colony formation,migration and invasion,compared with cells transfected with NC(P<0.05).Conclusion:RASAL1 is proved to be a target gene of miR-130b-5p and overexpression of miR-130b-5p results in promoted proliferation,colony formation,migration and invasion ability through targeted modulate of RASAL1.
Keywords/Search Tags:miR-130b-5p, RASAL1, Gastric cancer
PDF Full Text Request
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