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RNAi knockdown of pik3CA preferentially inhibits invasion of mutant pik3CA cells

World Journal of Gastroenterology 2011年第32期17卷 3700-3708页

作者： Xin-ke Zhou Sheng-Song Tang Gao Yi Min Hou Jin-Hui Chen Bo Yang Ji-Fang Liu Zhi-Min He Department of Pharmacology University of South China Hengyang 421001 Hunan Province China Institute of Cancer Research Affiliated Tumor Hospital of Guangzhou Medical College Guangzhou 510095 Guangdong Province China Laboratory of Applied Marine Biology Southem China Sea Institute of Oceanology Chinese Academy of Science Guangzhou 510300 Guangdong Province China

AIM: To explore the effects of siRNA silencing of pik3CA on proliferation, migration and invasion of gastric cancer cells and to investigate the underlying mechanisms. METHODS: The mutation of pik3CA in exons 9 and 20 of gastric cancer cell lines HGC-27, SGC-7901, BGC-823, MGC-803 and MkN-45 was screened by polymerase chain reaction (PCR) followed by sequencing. BGC-823 cells harboring no mutations in either of the exons, and HGC-27 cells containing pik3CA mutations were employed in the current study. siRNA targeting pik3CA was chemically synthesized and was transfect- ed into these two cell lines in vitro . mRNA and protein expression of pik3CA were detected by real-time PCR and Western blotting, respectively. We also measured phosphorylation of a serine/threonine protein kinase (akt) using Western blotting. The proliferation, migration and invasion of these cells were examined separately by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet-razolium bromide (MTT), wound healing and Transwell chambers assay. RESULTS: The siRNA directed against pik3CA effectively led to inhibition of both endogenous mRNA and protein expression of pik3CA, and thus significantly down-regulated phosphorylation of akt (P pik3CA resulted in an obvious reduction in tumor cell proliferation activity, migration and invasion potential (P 3 cells. knockdown of pik3CA in mutant HGC-27 cells contributed to a reduction in cell invasion to a greater extent than in non-mutant BGC-823 cells. CONCLUSION: siRNA mediated targeting of pik3CA may specifically knockdown the expression of pik3CA in gastric cancer cells, providing a potential implication for therapy of gastric cancer.

关键词： Gastric cancer Metastasis pik3CA pi3k/ akt pathway RNAi

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