Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:wangxingkun
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AIM: To study the effect of inhibited E-cadherin expression on invasion of cancer cells. METHODS: We designed the nucleotide sequence of siRNA corresponding to 5’ non-coding and coding sequence of E-cadherin. 21-nucleotide dssiRNA was synthesized by In vitro transcription with Ambion Silencer TM siRNA Construction Kit. siRNA was transfected into gastric cancer MKN45 using TransMessenger transfection Kit. RT-PCR and immunofluorescent assay were used to investigate the inhibition of the expression of mutated E-cadherin. Invasive ability of cancer cells was determined by Transwell assay. RESULTS: The synthesis of E-cadherin mRNA rather than protein expression was suppressed dramatically 7 d after interference. Decreased protein expression was observed on d 10 after interference. On d 11, invasion ability was enhanced significantly. CONCLUSION: siRNA targeted at non-coding and coding sequence of E-cadherin showed significant inhibition on mRNA and protein expression. Inhibited E-cadherin expression results in increased invasion ability of cancer cells. METHODS: We designed the nucleotide sequence of siRNA corresponding to 5 ’non-coding and coding sequence of E-cadherin. 21-nucleotide dssiRNA was synthesized by In Invitrogen ability of cancer cells was determined to inhibit the expression of mutated E-cadherin. by Transwell assay. RESULTS: The synthesis of E-cadherin mRNA rather than protein expression was suppressed dramatically for 7 d after interference. Decreased protein expression was observed on d 10 after interference. On d 11, invasion ability was significantly increased. CONCLUSION: siRNA targeted at non-coding and coding sequence of E-cadherin showed significant inhibition on mRNA and protein expression. Inhibited E-cadherin e xpression results in increased invasion ability of cancer cells.
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