CAJ | 학술논문

目的探讨人附睾蛋白4(HE4)基因表达下调对卵巢癌细胞增殖、黏附以及侵袭能力的影响.方法构建含HE4 shDNA的重组质粒,并将其转染卵巢癌细胞系SKOV3细胞,RNA干扰后,通过即时定量聚合酶链反应(qRT-PCR)、Western blot等测定HE4表达下调情况；同时利用平板克隆形成实验、细胞黏附实验、测定稳定转染细胞的侵袭力改变等方法,比较分析HE4相关的生物学效应.结果 Western blot以及qRT-PCR结果均提示HE4干扰重组质粒构建成功,稳定转染SKOV3细胞后能显著抑制HE4基因的表达.体外实验显示,HE4表达下调的细胞侵袭能力明显下降(P＜0.05),黏附率明显下降到27.3％,而增殖能力较转染前无明显变化.结论所构建的HE4干扰重组表达载体能有效抑制HE4基因表达,而HE4表达下调对卵巢癌的细胞黏附和侵袭能力降低.
목적 탐토인부고단백4(HE4)기인표체하조대란소암세포증식、점부이급침습능력적영향.방법 구건함HE4 shDNA적중조질립,병장기전염란소암세포계SKOV3세포,RNA간우후,통과즉시정량취합매련반응(qRT-PCR)、Western blot등측정HE4표체하조정황；동시이용평판극륭형성실험、세포점부실험、측정은정전염세포적침습력개변등방법,비교분석HE4상관적생물학효응.결과 Western blot이급qRT-PCR결과균제시HE4간우중조질립구건성공,은정전염SKOV3세포후능현저억제HE4기인적표체.체외실험현시,HE4표체하조적세포침습능력명현하강(P＜0.05),점부솔명현하강도27.3％,이증식능력교전염전무명현변화.결론 소구건적HE4간우중조표체재체능유효억제HE4기인표체,이HE4표체하조대란소암적세포점부화침습능력강저.
Objective To investigate the effects of HE4 gene knockdown on the proliferation,adhesion and invasion of the ovarian cancer cells SKOV3.Methods The knockdown of HE4 gene was performed by RNAi technology.The recombinant plasmids (pSUPER-HE4 shDN4s) were constructed and transfected into human ovarian cancer cells SKOV3.HE4 expression was then identified by real-time PCR and Western blot analysis.The invasion and adhesion ability of transduced cells were determined.In addition,cell proliferation and growth were analyzed by colonies formation assay.Results Knockdown of HE4 was achieved,and further confirmed by real-time PCR and Western blot.The proliferation of HE4-down-regulated cells was not affected,but the invasion ability of the transfected cells was reduced (P ＜ 0.05) and the adhesion ability was also reduced to 27.3％.Conclusion HE4 expression is down-regulated effectively by the constructed HE4 shDNA,and thus knockdown of HE4 inhibits the adhesion and invasion of SKOV3 cells.