CAJ | 학술논문

目的：探讨ER-α36基因与人雌激素受体（ ER）阴性乳腺癌细胞MDA-MB-231的内在联系。观察其对ER阴性乳腺癌细胞的影响。为探索ER阴性乳腺癌的基因治疗提供新的途径。方法利用RNAi技术，构建靶向ER-α36基因的短发夹状RNA（shRNA）重组质粒，脂质体法将pRNAT-U6.1/Neo-ER-α36导入雌激素受体阴性乳腺癌细胞MDA-MB-231，利用RT-PCR法检测MDA-MB-231细胞ER-α36基因表达的抑制情况；MTT法检测雌激素受体阴性乳腺癌细胞MDA-MB-231的增殖情况。结果①PCR、酶切鉴定、DNA测序证实小干扰质粒构建成功，无碱基突变；②ER-α36-siRNA表达载体转染ER阴性乳腺癌细胞MDA-MB-231可有效抑制ER-α36 mRNA表达（抑制效率＝86.3％），P＜0.01；③MTT实验结果表明ER-α36沉默可有效抑制ER阴性乳腺癌细胞MDA-MB-231的增殖，提示ER-α36参与MAPK/ERK信号转导通路调节细胞增殖。结论 ER-α36参与MAPK/ERK信号转导途径，与ER阴性乳腺癌的发生、发展关系密切。RNAi有效沉默ER-α36基因是ER阴性乳腺癌基因治疗的有效手段。ER-α36可能成为ER阴性乳腺癌基因治疗的新靶点。
목적：탐토ER-α36기인여인자격소수체（ ER）음성유선암세포MDA-MB-231적내재련계。관찰기대ER음성유선암세포적영향。위탐색ER음성유선암적기인치료제공신적도경。방법이용RNAi기술，구건파향ER-α36기인적단발협상RNA（shRNA）중조질립，지질체법장pRNAT-U6.1/Neo-ER-α36도입자격소수체음성유선암세포MDA-MB-231，이용RT-PCR법검측MDA-MB-231세포ER-α36기인표체적억제정황；MTT법검측자격소수체음성유선암세포MDA-MB-231적증식정황。결과①PCR、매절감정、DNA측서증실소간우질립구건성공，무감기돌변；②ER-α36-siRNA표체재체전염ER음성유선암세포MDA-MB-231가유효억제ER-α36 mRNA표체（억제효솔＝86.3％），P＜0.01；③MTT실험결과표명ER-α36침묵가유효억제ER음성유선암세포MDA-MB-231적증식，제시ER-α36삼여MAPK/ERK신호전도통로조절세포증식。결론 ER-α36삼여MAPK/ERK신호전도도경，여ER음성유선암적발생、발전관계밀절。RNAi유효침묵ER-α36기인시ER음성유선암기인치료적유효수단。ER-α36가능성위ER음성유선암기인치료적신파점。
Objectove To explore the intrinsic relation between ER-α36 and human ER-negative breast cancer cell MDA-MB-231,and observe its influences on ER-negative breast cancer cell. and offer new directions for ER-negative breast cancer treatment. Methods Construct the plasmid containing short hairpin RNA(ShRNA)of ER-α36. Sup-press the expression of ER-α36 gene by transfecting the plasmid into human ER-negative breast cancer cell lines MDA-MB-231. The recombinant vector was transfected into MDA-MB-231 cells with LipofectamineTM2000 and the ex-pression of ER-α36 was detected by RT-PCR. The proliferation of MDA-MB-231 were detected by MTT assay. Re-suIts ① DNA sequencing for the PCR product showed that the recombinant vector pRNAT-U6 . 1/Neo-ER-α36 was successfully constructed without any base pair mutation.②Transfection of MDA-MB-231 cells with ER-α36 siRNA plasmid significantly inhibited ER-α36 expression at mRNA level. The efficiency was up to 86. 3%(P<0. 01).③Silencing ER-α36 can inhibit the proliferation of human ER-negative breast cancer cell MDA-MB-231 . Result of MTT assay hint that ER-α36 is involved into the MAPK/ERK signaling pathway and stimulates cell growth. concIusoon ER-α36 is closer with the ER-negative breast cancer. The silence of ER-α36 by RNAi is an effective treatment in es-trogen receptor negative breast cancer. ER-α36 maybe the new target for molecular therapy of ER-negative breast cancer.