摘要目的 探究微小RNA-210(miR-210)在乳腺癌组织中的表达及临床意义,并探究其在体外对人三阴性乳腺癌细胞MDA-MB-231增殖及转移能力的影响.方法 收集湖南省肿瘤医院病理科2013年12月至2015年9月乳腺癌组织及相应癌旁组织标本各82例,荧光定量PCR(qRT-PCR)技术检测组织及细胞中miR-210的表达水平,分析miR-210与患者临床资料及预后的关系.三阴性乳腺癌细胞MDA-MB-231转染含miR-210全长的载体作为实验组,转染空白载体作为对照组,CCK-8实验检测两组细胞增殖能力,Transwell侵袭及迁移实验检测两组细胞转移及侵袭能力.结果 qRT-PCR结果显示乳腺癌组织中miR-210的表达水平为0.198±0.014,显著高于癌旁组织的0.084±0.009,差异具有统计学意义(t=8.141,P<0.001);三阴性乳腺癌组织miR-210表达水平为0.254±0.026,显著高于非三阴性乳腺癌组织0.167±0.015,差异具有统计学意义(t=3.175,P=0.003).miR-210高表达、低表达两组患者TNM分期、分子分型差异有统计学意义(χ2=7.859,P=0.005;χ2=7.053,P=0.008);而miR-210高表达的患者4年生存率显著低于低表达患者(49.37％:76.80％),差异具有统计学意义(χ2=4.743,P=0.024).qRT-PCR结果显示,实验组细胞miR-210的表达水平为0.517±0.038,显著高于对照组细胞的0.284±0.022,差异具有统计学意义(t=9.280,P<0.001).CCK-8实验结果显示实验组细胞48、72及96 h时增殖能力显著高于对照组细胞(3.771±0.452:3.206±0.314;7.662±0.619:6.736±0.552;15.477±1.425:11.592±1.243),差异具有统计学意义(t=2.296,P=0.025;t=2.496,P=0.019;t=4.594,P=0.001).Transwell侵袭实验结果显示实验组细胞下室面细胞数为(107.8±13.0)个,显著高于对照组细胞的(74.4±10.9)个,差异具有统计学意义(t=3.732,P=0.001);迁移实验结果显示实验组细胞下室面细胞数为(136.5±18.5)个,显著高于对照组细胞的(87.4±15.7)个,差异具有统计学意义(t=4.256,P<0.001).结论 miR-210在乳腺癌组织中高表达,且与患者病情进展、肿瘤恶性程度及预后密切相关,体外miR-210可促进三阴性乳腺癌MDA-MB-231细胞的恶性行为,是潜在的分子标志物及靶向治疗位点.

abstractsObjective To investigate the expression and clinical significance of microRNA-210 (miR-210)in breast cancer tissues,and to investigate its effect on the proliferation and metastasis of human triple negative breast cancer cell line MDA-MB-231 in vitro. Methods The breast cancer tissues and paracancerous tissues in 82 patients were collected in the Department of Pathology of Hunan Cancer Hospital from December 2013 to September 2015. Quantitative real-time polymerase chain reaction (qRT-PCR)technique was used to detect the expression level of miR-210 in tissues and cells. The relationship between the expression of miR-210 and clinical data and prognosis of patients were analyzed. The triple negative breast cancer cell line MDA-MB-231 transfected with full-length miR-210 plasmid was regarded as test group,and the cell transfected with blank vector was regarded as control group. CCK-8 assay was used to detect the proliferation ability of cells in both groups. Transwell invasion and migration assays were used to detect the metastasis and invasion ability of cells. Results The results of qRT-PCR showed that the expression level of miR-210 was 0. 198 ± 0. 014 in breast cancer tissues,which was significantly higher than that in paracancerous tissues (0. 084 ± 0. 009),and the difference was statistically significant (t = 8. 141,P < 0. 001). The expression level of miR-210 in triple nega-tive breast cancer tissues was 0. 254 ± 0. 026,which was significantly higher than that in non-triple negative breast cancer tissues (0. 167 ± 0. 015),and the difference was statistically significant (t = 3. 175,P =0. 003). There were significant differences in TNM staging and molecular typing between the patients with high and low expression of miR-210 (χ2 = 7. 859,P = 0. 005;χ2 = 7. 053,P = 0. 008). The 4-year survival rate of patients with high expression of miR-210 was significantly lower than that of patients with low expression of miR-210 (49. 37％ vs. 76. 80％),and the difference was statistically significant (χ2 = 4. 743,P = 0. 024). The results of qRT-PCR showed that the expression of miR-210 in cells in test group was 0. 517 ± 0. 038,which was significantly higher than that in control group (0. 284 ± 0. 022),and the difference was statistically significant (t = 9. 280,P < 0. 001). The results of CCK-8 assay showed that the proliferation abilities of the test group were significantly higher than those of the control group in 48,72 and 96 h (3. 771 ± 0. 452 vs. 3. 206 ± 0. 314;7. 662 ± 0. 619 vs. 6. 736 ± 0. 552;15. 477 ± 1. 425 vs. 11. 592 ± 1. 243),and the differences were statistically significant (t = 2. 296,P = 0. 025;t = 2. 496,P = 0. 019;t = 4. 594,P = 0. 001). The results of Transwell invasion assay showed that the cell number of test group in inferior surface was 107. 8 ± 13. 0,which was significantly higher than that of control group (74. 4 ± 10. 9),and the difference was statistically significant (t = 3. 732,P = 0. 001). The results of Transwell migration assay showed that the cell number of test group in inferior surface was 136. 5 ± 18. 5,which was significantly higher than that of control group (87. 4 ± 15. 7), and the difference was statistically significant (t = 4. 256,P < 0. 001). Conclusion The expression of miR-210 in breast cancer tissues is high,and its expression is closely related to progression,malignancy and progno-sis of patients. In vitro,miR-210 can promote the malignant behavior of triple negative breast cancer cell line MDA-MB-231. It is a potential molecular marker and targeted treatment site.