CAJ | 학술논문

　　背景与目的：miR-301在多种恶性肿瘤中表达上调，然而其在结肠癌中的表达及功能尚不清楚。本研究旨在检测微小RNA miR-301在结肠癌组织中的表达，并在体内外研究miR-301反义寡核苷酸技术(ASO)对结肠癌细胞增殖和凋亡的影响。方法：运用荧光定量PCR定量分析120例结肠癌患者癌组织及对应癌旁组织中miR-301的表达；通过miR-301ASO降低结肠癌SW620细胞中miR-301的表达，采用MTT、克隆形成实验、流式细胞技术及体内实验观察miR-301ASO对SW620细胞产生的生物学效应。结果：在120例结肠癌患者中，63.33%(76/120)的结肠癌组织miR-301表达明显高于对应癌旁组织(P=0.00)；与对照组miR-301的表达量(0.50±0.07)相比，miR-301ASO组可以显著降低miR-301的表达(0.09±0.01，P=0.00)；MTT实验结果显示，转染miR-301 ASO组24、48、96 h SW620细胞的存活数量均明显低于对照组(P<0.05)；克隆形成实验结果显示，miR-301ASO组克隆形成率(5.33%±0.74%)较对照组(33.33%±8.38%)显著降低(P=0.00)；体内研究进一步证实miR-301ASO可以抑制肿瘤细胞的增殖，从而导致肿瘤生长较对照组慢(P=0.01)，肿瘤的体积较对照组明显减小(P=0.01)；流式细胞仪检测显示，转染miR-301ASO组SW620细胞凋亡指数(15.68±1.46)较随机染转染ASO组(3.36±0.88)明显增高(P=0.02)；另外，降低miR-301的表达发现Bcl-2的mRNA和Bcl-2蛋白均明显下降(P=0.00，P=0.00)。结论：miR-301在结肠癌组织中表达上调，降低miR-301的表达可有效抑制结肠癌细胞生长、促进细胞凋亡。miR-301有可能成为结肠癌基因表达调控的新靶点。
　　배경여목적：miR-301재다충악성종류중표체상조，연이기재결장암중적표체급공능상불청초。본연구지재검측미소RNA miR-301재결장암조직중적표체，병재체내외연구miR-301반의과핵감산기술(ASO)대결장암세포증식화조망적영향。방법：운용형광정량PCR정량분석120례결장암환자암조직급대응암방조직중miR-301적표체；통과miR-301ASO강저결장암SW620세포중miR-301적표체，채용MTT、극륭형성실험、류식세포기술급체내실험관찰miR-301ASO대SW620세포산생적생물학효응。결과：재120례결장암환자중，63.33%(76/120)적결장암조직miR-301표체명현고우대응암방조직(P=0.00)；여대조조miR-301적표체량(0.50±0.07)상비，miR-301ASO조가이현저강저miR-301적표체(0.09±0.01，P=0.00)；MTT실험결과현시，전염miR-301 ASO조24、48、96 h SW620세포적존활수량균명현저우대조조(P<0.05)；극륭형성실험결과현시，miR-301ASO조극륭형성솔(5.33%±0.74%)교대조조(33.33%±8.38%)현저강저(P=0.00)；체내연구진일보증실miR-301ASO가이억제종류세포적증식，종이도치종류생장교대조조만(P=0.01)，종류적체적교대조조명현감소(P=0.01)；류식세포의검측현시，전염miR-301ASO조SW620세포조망지수(15.68±1.46)교수궤염전염ASO조(3.36±0.88)명현증고(P=0.02)；령외，강저miR-301적표체발현Bcl-2적mRNA화Bcl-2단백균명현하강(P=0.00，P=0.00)。결론：miR-301재결장암조직중표체상조，강저miR-301적표체가유효억제결장암세포생장、촉진세포조망。miR-301유가능성위결장암기인표체조공적신파점。
Background and purpose: The miR-224 in a variety of malignant tumors is overexpression, however, its expression and function in colon cancer are not clear. The aim of this study was to investigate the expression of miR-301 in colon cancer tissues and demonstrate the regulative effects of miR-301 ASO on the proliferation and apoptosis of colon cancer cell in vitro and in vitro. Methods:The expression of miR-301 in 120 colon cancer tissues and their adjacent tissues was detected by real-time quantitative PCR method. After transfection with miR-301ASO, the biological effects of miR-301 in SW620 cells were measured by MTT assay, the colony formation experiment, flow cytometry and the in vivo experiment. Results: The expression level of miR-301 was found to be overexpressed in 63.33% (76/120) of the colon cancer cases (P<0.05). miR-301 expression in SW620 cells (transfection with miR-301 ASO, 0.09±0.01) was significantly less than control group (0.50±0.07, P=0.00). MTT assay results showed that SW620 cells survived rate at 24, 48 and 96 h decreased greatly after transfection with miR-301ASO (P=0.00). Clone formation assay revealed that miR-301 ASO group colony formation rate (5.33%±0.74%) was significantly lower than the control group (33.33%±8.38%, P=0.00). In vivo study further confirmed that miR-301ASO could inhibit the proliferation of SW620 cells (P<0.05), and miR-301ASO group grew substantially slow compared with the negative control group (P=0.00). Flow cytometry indicated that the apoptotic index in miR-301 ASO group (15.68±1.46) was significantly higher than the control group (3.36±0.88, P=0.02). In addition, the Bcl2 mRNA and protein were significantly decreased after reduce the expression of miR-301 (P=0.00, P=0.00). Conclusion:MiR-301 was overexpressed in human colon cancer. Reduce the expression of miR-301 can effectively inhibit the growth of colon cancer cells and promote apoptosis. MiR-301 may become a new target for the regulation of gene expression in colon cancer.