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Effect of P115 shRNA on Angiogenesis of Gastric Carcinoma in Vitro

Author: WenXue
Tutor: YiYongFen
School: Chongqing Medical University
Course: Pathology and Pathophysiology
Keywords: p115 MIF HUVEC angiogenesis
CLC: R735.2
Type: Master's thesis
Year: 2011
Downloads: 21
Quote: 1
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Abstract


ObjectiveTo investigate the effect of p115shRNA in gastric cancer cell lines on angiogenesis of HUVEC and its mechanism.MethodsThe BGC-823 cells were divided into untransfected group, empty vector control group, and p115shRNA-1318 transfected group. The BGC-823 cells stably expressing p115 gene were established by G418.The expression of p115, MIF, p-Akt and VEGF-A in gastric cancer cells was measured by Western blot analysis. The p115, MIF and VEGF-A mRNA levels were detected by reverse transcript-polymerase chain reaction(RT-PCR). Expressions of p115, MIF and VEGF-A were analyzed by immunofluorescence cytochemistry. Migration and capillary-like structures formation of HUVEC were assessed by transwell migration assay and matrigel assay, respectively. The effect of p115shRNA on HUVEC proliferation was analyzed with cell counting kit-8 (CCK-8).ResultsThe results of Western blot showed that the expression levels of p115, MIF, p-Akt and VEGF-A were reduced in p115 shRNA BGC823 cells (P<0.05). The p115 shRNA significantly inhibited the expressions of p115, MIF and VEGF-A. Immunofluorescence showed that their inhibitory rate were 63%,65%,68%, respectively. HUVEC migration in p115-silencing BGC823 cell lines(39±5) were significantly weaker than that in BGC823 cells of untransfected group(174±4) and empty vector control group (179±4)(P < 0.05). Compared with the other two groups, the proliferation of HUVEC was obviously lower (P < 0.05). The formation of HUVEC in p115-silencing cells(422±41) was significantly weaker than that in the other two groups (1289±37,1329±33)(P < 0.05). ConclusionThese studies demonstrate that p115shRNA can reduce MIF and VEGF-A expressions and may suppress angiogenesis of HUVEC in gastric cancer via activation of PI3K/Akt signal pathway.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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