山西医科大学学报
山西醫科大學學報
산서의과대학학보
Journal of Shanxi Medical University
2015年
10期
977-981
,共5页
吕娟%李婧%张春梅%郭冬丽%王赞宏
呂娟%李婧%張春梅%郭鼕麗%王讚宏
려연%리청%장춘매%곽동려%왕찬굉
Derlin-1%宫颈癌%HeLa%RNA干扰
Derlin-1%宮頸癌%HeLa%RNA榦擾
Derlin-1%궁경암%HeLa%RNA간우
Derlin-1%cervical cancer%HeLa cells%RNA interference
目的 采用RNAi技术抑制Derlin-1基因的表达,探讨其表达降低后对宫颈癌HeLa细胞生长的影响. 方法 合成针对Derlin-1基因的干扰片段,构建重组质粒,用脂质体法将重组质粒转染人宫颈癌HeLa细胞,并筛选出稳定表达株,作为siRNA-Derlin-1组,选取空质粒转染后的HeLa细胞为空质粒组,未做任何处理的HeLa细胞为空白对照组,采用RT-PCR检测重组质粒对HeLa细胞Derlin-1 mRNA的影响,Western blot检测其对Derlin-1蛋白表达的影响,MTT法分析其对细胞增殖的影响,ELISA法检测其对细胞凋亡的影响,同时检测凋亡因子caspase-3的表达. 结果 与空质粒组和空白对照组相比,siR-NA-Derlin-1组HeLa细胞中Derlin-1 mRNA和蛋白的表达显著降低(P<0. 05),MTT法显示siRNA-Derlin-1组宫颈癌HeLa细胞增殖能力显著低于空质粒组和空白对照组(P<0. 05),同时siRNA-Derlin-1组凋亡细胞增加,凋亡核心蛋白caspase-3表达增加. 结论 抑制内质网相关降解蛋白Derlin-1的表达能有效降低宫颈癌HeLa细胞的增殖,促进凋亡.
目的 採用RNAi技術抑製Derlin-1基因的錶達,探討其錶達降低後對宮頸癌HeLa細胞生長的影響. 方法 閤成針對Derlin-1基因的榦擾片段,構建重組質粒,用脂質體法將重組質粒轉染人宮頸癌HeLa細胞,併篩選齣穩定錶達株,作為siRNA-Derlin-1組,選取空質粒轉染後的HeLa細胞為空質粒組,未做任何處理的HeLa細胞為空白對照組,採用RT-PCR檢測重組質粒對HeLa細胞Derlin-1 mRNA的影響,Western blot檢測其對Derlin-1蛋白錶達的影響,MTT法分析其對細胞增殖的影響,ELISA法檢測其對細胞凋亡的影響,同時檢測凋亡因子caspase-3的錶達. 結果 與空質粒組和空白對照組相比,siR-NA-Derlin-1組HeLa細胞中Derlin-1 mRNA和蛋白的錶達顯著降低(P<0. 05),MTT法顯示siRNA-Derlin-1組宮頸癌HeLa細胞增殖能力顯著低于空質粒組和空白對照組(P<0. 05),同時siRNA-Derlin-1組凋亡細胞增加,凋亡覈心蛋白caspase-3錶達增加. 結論 抑製內質網相關降解蛋白Derlin-1的錶達能有效降低宮頸癌HeLa細胞的增殖,促進凋亡.
목적 채용RNAi기술억제Derlin-1기인적표체,탐토기표체강저후대궁경암HeLa세포생장적영향. 방법 합성침대Derlin-1기인적간우편단,구건중조질립,용지질체법장중조질립전염인궁경암HeLa세포,병사선출은정표체주,작위siRNA-Derlin-1조,선취공질립전염후적HeLa세포위공질립조,미주임하처리적HeLa세포위공백대조조,채용RT-PCR검측중조질립대HeLa세포Derlin-1 mRNA적영향,Western blot검측기대Derlin-1단백표체적영향,MTT법분석기대세포증식적영향,ELISA법검측기대세포조망적영향,동시검측조망인자caspase-3적표체. 결과 여공질립조화공백대조조상비,siR-NA-Derlin-1조HeLa세포중Derlin-1 mRNA화단백적표체현저강저(P<0. 05),MTT법현시siRNA-Derlin-1조궁경암HeLa세포증식능력현저저우공질립조화공백대조조(P<0. 05),동시siRNA-Derlin-1조조망세포증가,조망핵심단백caspase-3표체증가. 결론 억제내질망상관강해단백Derlin-1적표체능유효강저궁경암HeLa세포적증식,촉진조망.
Objective To investigate the effect of Derlin-1 gene inhibition mediated by RNA interference on the proliferation of cervi-cal cancer HeLa cells in vitro. Methods Recombinant plasmids for RNA interference of Derlin-1 gene were constructed and transfect-ed into HeLa cells via lipofectamine 2000, which were named as siRNA-Derlin-1 group. Meanwhile, HeLa cells transfected with empty plasmids were chosen as empty plasmid group, and the untreated HeLa cells were selected as blank control group. The mRNA and pro-tein expression of Derlin-1 in the stably transfected cells was detected by RT-PCR and Western blot. The changes of cell proliferation were detected by MTT assay. Enzyme-linked immunosorbent assay( ELISA) was employed to observe the effect of transfection on the apoptosis. The expression of caspase-3 in the transfected cells was also detected by RT-PCR and Western blot. Results Compared with empty plasmids group and blank control group, the mRNA and protein expression levels of the HeLa cells in Derlin-1-siRNA group were significantly decreased(P<0. 05),and the cell proliferation was inhibited(P<0. 05). The apoptosis cells were increased in Der-lin-1-siRNA group and the expression of caspase-3 was effectively promoted compared with empty plasmids group and blank control group(P<0. 05). Conclusion Derlin-1 gene could be highly expressed in cervical cancer and RNA interference-mediated Derlin-1 silencing can strongly inhibit the proliferation and induce apoptosis in HeLa cells.
