【摘要】：目的：探討HSP90α在食管癌細(xì)胞侵襲轉(zhuǎn)移中的作用機(jī)制。方法：使用細(xì)胞轉(zhuǎn)染技術(shù)將siRNA-HSP90α轉(zhuǎn)染入CE81T-4細(xì)胞中,并分為實(shí)驗(yàn)組、陰性對(duì)照組、空白對(duì)照組;并用RT-PCR和Western blot檢測(cè)3組細(xì)胞中HSP90αmRNA水平和蛋白質(zhì)水平的表達(dá)差異,驗(yàn)證是否轉(zhuǎn)染成功。MTT法檢測(cè)3組細(xì)胞的增值活力,流式細(xì)胞術(shù)分析細(xì)胞周期、凋亡的改變,劃痕實(shí)驗(yàn)和Transwell體外侵襲實(shí)驗(yàn)檢測(cè)轉(zhuǎn)染后CE81T-4細(xì)胞遷移和侵襲的變化。結(jié)果：在siRNA-HSP90α成功轉(zhuǎn)染CE81T-4細(xì)胞后,RT-PCR檢測(cè)3組結(jié)果顯示,實(shí)驗(yàn)組中HSP90αmRNA水平表達(dá)量顯著低于空白對(duì)照組和陰性對(duì)照組(P 0.05, P 0.05)。Western blot分析顯示實(shí)驗(yàn)組HSP90α蛋白表達(dá)量降低,與空白對(duì)照組和陰性組相比差異有統(tǒng)計(jì)學(xué)意義(P0.05,P0.05)。MTT實(shí)驗(yàn)結(jié)果顯示,實(shí)驗(yàn)組細(xì)胞的增殖能力明顯減弱,與其他兩組比較差異具有統(tǒng)計(jì)學(xué)意義(P0.05,P0.05)。HSP90a基因表達(dá)沉默后,細(xì)胞凋亡率為32.67%,較空白對(duì)照組和陰性對(duì)照組明顯增加(P0.05,P0.05)。實(shí)驗(yàn)組細(xì)胞周期被阻滯在G0/G1期。Transwell體外侵襲實(shí)驗(yàn)顯示實(shí)驗(yàn)組侵襲遷移的細(xì)胞個(gè)數(shù)為(76.4±8.7),顯著低于空白照組(140.3±5.8,P0.05)與陰性對(duì)照組(129.3±10.1,P0.05)。劃痕實(shí)驗(yàn)結(jié)果顯示,72h后實(shí)驗(yàn)組細(xì)胞的遷移力較其他兩組明顯降低。結(jié)論：HSP90a的下調(diào)會(huì)降低食管癌細(xì)胞的轉(zhuǎn)移侵襲能力。
[Abstract]:Objective: to investigate the role of HSP90 偽 in invasion and metastasis of esophageal cancer cells. Methods: siRNA-HSP90 偽 was transfected into CE81T-4 cells by cell transfection technique, and was divided into experimental group, negative control group and blank control group, and the expression of HSP90 偽 mRNA and protein in the three groups were detected by RT-PCR and Western blot. The cell cycle and apoptosis were analyzed by flow cytometry, and the changes of migration and invasion of CE81T-4 cells after transfection were detected by scratch test and Transwell invasion assay in vitro. Results: after siRNA-HSP90 偽 was successfully transfected into CE81T-4 cells, the results of RT-PCR showed that the expression of HSP90 偽 mRNA in the experimental group was significantly lower than that in the blank control group and the negative control group (P 0.05, P 0.05). Western blot analysis showed that the expression of HSP90 偽 protein in the experimental group was significantly lower than that in the control group. Compared with the blank control group and the negative group, the difference was statistically significant (P 0.05 P 0.05). MTT assay showed that the proliferation ability of the experimental group was significantly decreased, and the difference was statistically significant compared with the other two groups (P 0.05 P 0.05) .HSP90a gene expression silenced. The apoptosis rate was 32.67%, which was significantly higher than that in the blank control group and the negative control group (P 0.05). The cell cycle of the experimental group was blocked in the G0/G1 phase. Transwell invasion in vitro showed that the number of the cells in the experimental group was (76.4 鹵8.7), significantly lower than that in the blank irradiation group (140.3 鹵5.8) and the negative control group (129.3 鹵10.1). The results of scratch test showed that the migration ability of the experimental group was significantly lower than that of the other two groups after 72 hours. Conclusion the down-regulation of% HSP90a may decrease the metastasis and invasion ability of esophageal cancer cells.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.1
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本文編號(hào)：2174990