CAJ | 학술논문

目的：研究辛伐他汀（simvastatin，SIM）对人乳腺癌MCF-7细胞内多能干细胞标志物Oct3/4、Nanog和Sox-2表达的影响。方法：应用实时荧光定量聚合酶链式反应（qRT-PCR）技术、免疫荧光染色方法，流式细胞仪和免疫印迹（Western blot）方法检测SIM对MCF-7细胞内多能干细胞标志物表达的影响。结果：qRT-PCR结果显示，10、50和100μmol/L SIM作用于MCF-7细胞48h后，能显著抑制细胞内Oct3/4、Nanog和Sox-2基因表达，与对照组相比，差异有统计学意义（P<0.05），而SIM 1μmol/L浓度组和对照组相比差异无统计学意义（P>0.05）。SIM 50、100μmol/L浓度组和10μmol/L浓度组相比，抑制Oct3/4和Nanog的表达差异有统计学意义（P<0.05），而对Sox-2的表达抑制，SIM 10、50μmol/L和100μmol/L各浓度组间差异无统计学意义（P>0.05）。免疫荧光染色显示经10μmol/L SIM处理48 h后MCF-7细胞核内Oct3/4、Nanog和Sox-2蛋白表达减弱，部分细胞核无表达。流式细胞检测显示MCF-7细胞经10μmol/L SIM处理48 h后，Oct3/4阳性细胞数、Nanog阳性细胞数和Sox-2阳性细胞数显著减少（P<0.05），Western blot进一步证实经10μmol/L SIM处理48 h后MCF-7细胞核内Oct3/4、Nanog和Sox-2蛋白表达显著减少（P<0.05）。结论：SIM在体外能有效地抑制人乳腺癌MCF-7细胞内多能干细胞标志物Oct3/4、Nanog和Sox-2的表达，为SIM应用于癌症治疗提供实验依据。
목적：연구신벌타정（simvastatin，SIM）대인유선암MCF-7세포내다능간세포표지물Oct3/4、Nanog화Sox-2표체적영향。방법：응용실시형광정량취합매련식반응（qRT-PCR）기술、면역형광염색방법，류식세포의화면역인적（Western blot）방법검측SIM대MCF-7세포내다능간세포표지물표체적영향。결과：qRT-PCR결과현시，10、50화100μmol/L SIM작용우MCF-7세포48h후，능현저억제세포내Oct3/4、Nanog화Sox-2기인표체，여대조조상비，차이유통계학의의（P<0.05），이SIM 1μmol/L농도조화대조조상비차이무통계학의의（P>0.05）。SIM 50、100μmol/L농도조화10μmol/L농도조상비，억제Oct3/4화Nanog적표체차이유통계학의의（P<0.05），이대Sox-2적표체억제，SIM 10、50μmol/L화100μmol/L각농도조간차이무통계학의의（P>0.05）。면역형광염색현시경10μmol/L SIM처리48 h후MCF-7세포핵내Oct3/4、Nanog화Sox-2단백표체감약，부분세포핵무표체。류식세포검측현시MCF-7세포경10μmol/L SIM처리48 h후，Oct3/4양성세포수、Nanog양성세포수화Sox-2양성세포수현저감소（P<0.05），Western blot진일보증실경10μmol/L SIM처리48 h후MCF-7세포핵내Oct3/4、Nanog화Sox-2단백표체현저감소（P<0.05）。결론：SIM재체외능유효지억제인유선암MCF-7세포내다능간세포표지물Oct3/4、Nanog화Sox-2적표체，위SIM응용우암증치료제공실험의거。
Objective:To investigate the effect of simvastatin on expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells. Methods:Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunofluo-rescent staining, flow cytometry, and Western blot were used to detect the expression of pluripotency markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells treated with different doses of simvastatin. Results:qRT-PCR revealed the more signifi-cant inhibition of gene expressions of Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells when subjected to high doses of simvastatin (10, 50, and 100 μmol/L) compared with the control group (P<0.05). By contrast, no significant difference was observed be-tween the expressions of low-dose simvastatin treatment (1 μmol/L) and control (P>0.05). The inhibitory effect of simvastatin on the gene expressions of Oct3/4 and Nanog was more significantly apparent at 50 and 100 μmol/L dosages than at 10 μmol/L (P<0.05). By contrast, no significant difference in the inhibitory expression of Sox-2 was observed among the three high-dose treatments (P>0.05). Between the two higher-dose treatments (50 and 100 μmol/L), no significant difference in the inhibitory expressions of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was found. Meanwhile, in the 10 μmol/L simvastatin treatment, immunoflurescent staining showed a marked reduction in the protein expression of all three pluripotent markers in MCF-7 cells, and flow cytometry demonstrated a decrease of Oct3/4-, Nanog-, and Sox-2-positive cells (P<0.05). Western blot further revealed that the protein expression of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was significantly declined by the same simvastatin dose (P<0.05). Conclusion: Simvastatin can inhibit the expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells, proving the anti-cancer properties of simvastatin.