CAJ | 학술논문

目的探讨结肠癌LS174T细胞及结直肠腺癌组织中缺氧诱导因子-1α(HIF-1α)的表达及与Survivin的关系.方法构建靶向HIF-1α的质粒pSilence-2.1-U6-siRNA并鉴定,采用阳离子脂质体转染法将其转染LS174T细胞,低氧培养24h,应用逆转录-聚合酶链反应(RT-PCR)、Western blot检测siRNA对HIF-1α的抑制作用.Western blot检测Survivin蛋白表达.应用免疫组织化学方法检测69例人结直肠腺癌组织中HIF-1α、Survivin蛋白的表达.结果 LS174T细胞转染靶向HIF-1α的siRNA表达载体后,HIF-1α、Survivin表达显著下降.69例结直肠腺癌组织中,HIF-1α、Survivin蛋白的阳性表达率为66.7％ (46/69)和56.5％ (39/69).腺癌组HIF-1α蛋白阳性表达率显著高于腺瘤组(P＜0.01).Ⅲ期腺癌HIF-1α蛋白表达显著高于Ⅰ～Ⅱ患者(P＜0.05).结直肠腺癌中HIF-1α表达与Survivin显著正相关.结论结直肠腺癌发展过程中HIF-1α与Survivin表达密切相关.
목적 탐토결장암LS174T세포급결직장선암조직중결양유도인자-1α(HIF-1α)적표체급여Survivin적관계.방법 구건파향HIF-1α적질립pSilence-2.1-U6-siRNA병감정,채용양리자지질체전염법장기전염LS174T세포,저양배양24h,응용역전록-취합매련반응(RT-PCR)、Western blot검측siRNA대HIF-1α적억제작용.Western blot검측Survivin단백표체.응용면역조직화학방법검측69례인결직장선암조직중HIF-1α、Survivin단백적표체.결과 LS174T세포전염파향HIF-1α적siRNA표체재체후,HIF-1α、Survivin표체현저하강.69례결직장선암조직중,HIF-1α、Survivin단백적양성표체솔위66.7％ (46/69)화56.5％ (39/69).선암조HIF-1α단백양성표체솔현저고우선류조(P＜0.01).Ⅲ기선암HIF-1α단백표체현저고우Ⅰ～Ⅱ환자(P＜0.05).결직장선암중HIF-1α표체여Survivin현저정상관.결론 결직장선암발전과정중HIF-1α여Survivin표체밀절상관.
Objective To investigate the correlation between the expression levels of hypoxia inducible factor-1α (HIF-1α) and Survivin in human the colorectal carcinomas (CRCs).Methods Plasmid expressing small interfering RNA (siRNA) targeting HIF-1 α mRNA (pSilence-2.1-U6-siRNA ) was constructed and transfected into LS174T cells by Lipofectamine methods.The effect of HIF-1α on the expression of survivin by siRNA targeting HIF-1α mRNA was studied.The expression of survivin and HIF-1α was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.The expression of HIF-1 α and Survivin was investigated by immunohistochemistry in 69 patients with CRCs.Results The successful construction of pSilence-2.1-U6-siRNA plasmid was confirmed by sequencing analysis.HIF-1 α and Survivin expression levels were markedly down-regulated after the siRNA expression vector targeting HIF-1α was transfected into the LS174T cells.The positive rate of HIF-1α protein in CRCs was significantly higher than that in colorectal adenoma lesions ( P ＜ 0.05 ).There was significant difference in the expression of HIF-1α protein between stage Ⅲ and stage Ⅰ-Ⅱ CRCs (P ＜0.01 ).The level of HIF-1αwas positively correlated with that of Survivin protein ( P ＜ 0.05 ) during colorectal tumor progression.Conclusion There is a positive correlation between the expression levels of HIF-1αt and Survivin in tumor progression of human CRCs.