人附睾蛋白4基因表达下调对卵巢癌细胞生物学行为的影响

Effect of down-regulation of HE4 gene expression on biologic behavior of ovarian cancer cells

目的探讨人附睾蛋白4（HFA）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HEdshDNA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HFA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HFA干扰重组质粒构建成功，稳定转染SKOV3细胞后能显著抑制HFA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HFA干扰重组表达载体能有效抑制HFA基因表达，而HFA表达下调对卵巢癌的细胞黏附和侵袭能力降低。

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 shDNAs） 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.