Effect Of MiR-185 To The Invasion And Metastasis Of Gastric Cancer Cell SGC-7901 And The Expression Of CDC42

Objective:Explore the function of mi R-185 in the invasion and metastasis of gastric carcinoma cell SGC-7901,further illuminate the influence of mi R-185 to the target gene of CDC42.Methods: 1. By using the result of prophase mi RNA microarray chip and select the purpose mi RNA, the q RT- PCR technology further validates the result of the chips. 2. Mi R-185 or nonsense sequence was transfected into cell SGC-7901 respectively with the method of transient transfection, the condition of cell transfection was observed by inverted fluorescence microscope, transfection efficiency was further tested by q RT-PCR. The influence of invasion and metastasis after gastric carcinoma cells transfected was tested by Trans-well chamber method and scratch test. The target gene of purpose mi RNA was predicted by biological information software, q RT-PCR detected the gene of CDC42 m RNA trascripition expression levels after transfection, western blot method detected the protein expression levels.Result: 1. q RT- PCR to validate the result for the gene chips of the alternative mi RNA. 2. Select the mi R-185 as research objective, using biological information software to select the CDC42 as its target gene. After 24 hours of gastric cancer cell SGC- 7901 transfection, Lipofectamine3000 for transfection reagent in SGC-7901, the transfection efficiency is around 80% was tested by inverted fluorescence microscope. After transfected with mi R-185 mimics, q RT-PCR indicates that mi R-185 was increased significantly in SGC-7901. The Trans-well chamber method and scratch test results demonstrated that mi R-185 transfected cells had a decrease on the invasion and metastasis ability. QRT-PCR showed that the m RNA expression level of CDC42 in mi R-185 transfected cells were obviously decreased, and moreover, Western bolt results showed that the protein expression level of CDC42 in mi R-185 transfected cells were obviously decreased compared with transfection reagent group, untreated control group or the negative group.Conclusions: 1. Overexpression of mi R-185 might inhibit the invasion and metastasis of gastric cancer cell SGC-7901. 2. Overexpression of mi R-185 might down-regulate the m RNA and protein of CDC42, mi R-185 might through the gene of CDC42 play an important role in gastric cancer. 3. Mi R- 185 may play a role of tumor suppressor genes in the progress of gastric cancer.