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siRNA干扰即刻早期基因Egr-1对汉防己甲素诱导的胰腺癌细胞自噬、凋亡的作用研究

Keywords: 胰腺癌, 汉防己甲素, Egr-1, 细胞自噬, 细胞凋亡
pancreaticcancer
, tetrandrine, Egr-1, autophagy, cellapoptosis

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Abstract:

本实验将胰腺癌细胞PaTu8988分别转染无关干扰对照组(siRNA-Scramble)和干扰即刻早期基因Egr-1组(siRNA-Egr-1),再用不同浓度的汉防己甲素处理24h后,CCK-8法检测不同浓度的汉防己甲素对细胞活力的影响;Westernblot检测细胞自噬和凋亡相关蛋白表达水平的改变.结果表明:(1)低浓度的汉防己甲素能显著激活PaTu8988细胞自噬,但对增殖和凋亡无明显影响;(2)与对照组相比,转染了siRNA-Egr-1组自噬相关蛋白Beclin-1表达量显著下降,LC3II/LC3I比值下调;凋亡相关蛋白Bcl2/Bax的比值降低.上述结果表明,干扰即刻早期基因Egr-1的表达可以有效抑制低浓度汉防己甲素引起的胰腺癌细胞PaTu8988自噬,促进细胞凋亡.
HumanpancreaticcancercellsPaTu8988weretransfectedwithsiRNA-ScrambleandsiRNA-Egr-1for24hrespectively,thentreatedwithtetrandrineforanother24h.Treatedbydifferentconcentrationoftetrandrine,thecytotoxiceffectoftetrandrinewasexaminedbyCCK-8,andexpressionofautophagy-relatedproteinweredetectedbyWesternblot.Theexpressionofcellautophagy-andapoptosis-relatedproteinweredetectedbyWesternblot.Alloftheresultsindicatedthatcomparingwithcontrolgroup,siEgr-1significantlyattenuatestetrandrine-inducedautophagyandpromotestheapoptosisofPaTu8988cells

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