CAJ | 학술논문

目的应用RNA干扰术靶向降低乳腺癌MCF-7细胞中骨桥蛋白基因(OPN)的表达水平,研究OPN基因对乳腺癌细胞增殖和侵袭性等生物学行为的影响.方法将设计合成的骨桥蛋白OPN序列特异性小分子干扰RNA(siRNA),转染乳腺癌细胞株MCF-7,用逆转录PCR和蛋白印记杂交测OPN mRNA及蛋白水平的表达,通过生长曲线、克隆形成实验来检测细胞增殖及侵袭性情况的变化.结果 OPN 特异性siRNA转染后,骨桥蛋白mRNA及蛋白水平均明显下降,比色法显示转染组72 h吸光度值为(0.21±0.06)与空白对照组(1.18 ±0.05)相比较明显受到抑制.转染组细胞克隆形成数为3.8个,较空白对照组(7.3个)及阴性对照组(7.1个)明显下降(P<0.05).结论乳腺癌MCF-7细胞中,针对OPN合成的siRNA能够有效地抑制骨桥蛋白的表达,从而显著抑制细胞的增殖和侵袭能力.
목적 응용RNA간우술파향강저유선암MCF-7세포중골교단백기인(OPN)적표체수평,연구OPN기인대유선암세포증식화침습성등생물학행위적영향.방법 장설계합성적골교단백OPN서렬특이성소분자간우RNA(siRNA),전염유선암세포주MCF-7,용역전록PCR화단백인기잡교측OPN mRNA급단백수평적표체,통과생장곡선、극륭형성실험래검측세포증식급침습성정황적변화.결과 OPN 특이성siRNA전염후,골교단백mRNA급단백수평균명현하강,비색법현시전염조72 h흡광도치위(0.21±0.06)여공백대조조(1.18 ±0.05)상비교명현수도억제.전염조세포극륭형성수위3.8개,교공백대조조(7.3개)급음성대조조(7.1개)명현하강(P<0.05).결론 유선암MCF-7세포중,침대OPN합성적siRNA능구유효지억제골교단백적표체,종이현저억제세포적증식화침습능력.
Objective To study the effect of osteopontin (OPN) expression down-regulated by RNA inter-ference(RNAi) on the proliferation and invasion of breast carcinoma cell line MCF-7. Methods MCF-7 cells were transfected with the chemically synthesized small interfering RNA (siRNA). RT-PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The proliferation and invasion of MCF-7 were observed by cell growth curve and colony formation, experiments. Results After being transfected by sequence-specific siRNA targe-ting OPN, the expression of OPN RNA and protein decreased in MCF-7 cells. MTT test showed that the proliferation was inhibited significantly by siRNA compared with the control groups. The number of formed colonies was decreas ed in MCF-7 cells transfected with OPN - specific siRNA compared with control groups (P < 0.05). Condusion Sequence-specific siRNA targeting OPN can effectively inhibit the expression of OPN mRNA and protein levels, and significantly diminishes the proliferation and invasion of MCF-7 cells.