WWOX基因过表达对人胃癌细胞增殖、凋亡能力的影响及机制研究

Study on the effect and mechanism of overexpression of WWOX gene on proliferation and apoptosis of human gastric cancer cells

目的探究含有WW结构域的氧化还原酶基因(WWOX)mRNA在胃癌中的表达量及过表达WWOX对人胃癌SGC7901细胞增殖、凋亡能力的影响及作用机制。方法使用实时荧光定量PCR(qRT-PCR)检测WWOX mRNA在胃癌组织和癌旁组织中的表达量;利用脂质体LipofectamineTM2000将重组质粒pc DNA3.0-WWOX、空载体pc DNA3.0-NC转入人胃癌SGC7901细胞中,未转染细胞为空白组,蛋白质印迹法(Western blotting)检测WWOX蛋白表达量;绘制转染后人胃癌SGC7901细胞增殖状况;流式细胞术检测SGC7901细胞凋亡率;Western blotting检测转染后细胞周期蛋白D1(Cyclin D1)和生存素(Survivn)蛋白的表达量。结果 WWOX mRNA在胃癌组织中的表达量显著低于癌旁组织(P<0.05);pc DNA3.0-WWOX组WWOX蛋白的表达量显著高于pc DNA3.0-NC组和空白组(P<0.05);绘制细胞生长曲线发现,WWOX基因能抑制胃癌SGC7901细胞的增殖;流式细胞术检测结果显示,pc DNA3.0-WWOX组细胞的凋亡率显著高于pc DNA3.0-NC组和空白组(P<0.05);pc DNA3.0-WWOX组细胞Cyclin D1蛋白的表达显著低于pc DNA3.0-NC组和空白组(P<0.05),而Survivn蛋白显著高于pc DNA3.0-NC组和空白组(P<0.05)。结论 WWOX mRNA在胃癌中的表达量低于癌旁组织;过表达WWOX可通过降低Cyclin D1蛋白、增加Survivn蛋白抑制胃癌SGC7901细胞的增殖,并促进其凋亡。

Objective To investigate the expression of WWOX mRNA in gastric cancer and the effect of proliferation and apoptosis of up-regulation the expression of WWOX. Methods The expression of WWOX mRNA in gastric cancer and matched tumor-adjacent tissues was detected by qRT-PCR. The recombinant plasmid pcDNA3.0-WWOX and empty vector pcDNA3.0-NC were transfected into SGC7901 cell, and the expression of WWOX protein was detected by Western blotting, cell proliferation was counted and apoptosis was observed by flow cytometry. Results The expression of WWOX mRNA in gastric cancer was significantly lower than that in matched tumor-adjacent tissues （P 〈 0.05）. The expression of WWOX protein in the pcDNA3. 0-WWOX experimental group was obviously higher than that in the pcDNA3.0-NC control group and blank group （P 〈 0.05）. The proliferation rate of SGC7901 cells was lower and apoptosis rate of SGC7901 ceils was higher in pcDNA3.0-WWOX group than those in pcDNA3.0-NC control group and blank group （P 〈 0.05）. The level of Cyclin D1 protein in the pcDNA3.0-WWOX experimental group was obviously lower than that in the pcDNA3.0-NC control group and blank group （P 〈 0.05 ）, but Survivn protein was opposite. Conclusion WWOX gene shows low expression in gastric cancer. Up-regulation of WWOX can noticeably inhibit the proliferation and pro- mote apoptosis in SGC7901 ceils by reducing the Cyclin D1 protein and increasing the Survivn protein.