| 刘春梅,齐学林,马亚萍,杨亚峰,张秀德,潘 娟,王永席,宗 伟,姜孝新.miR-221对甲状腺乳头癌生物学特性的影响[J].现代生物医学进展英文版,2017,17(18):3438-3442. |
| miR-221对甲状腺乳头癌生物学特性的影响 |
| Effects of miR-221 on the Biological Characteristics of Papillary Thyroid Carcinoma |
| Received:November 01, 2016 Revised:November 26, 2016 |
| DOI：10.13241/j.cnki.pmb.2017.18.008 |
| 中文关键词: miR-221 甲状腺乳头癌 增殖 迁移 凋亡 |
| 英文关键词: miR-221 Papillary thyroid carcinoma Proliferation Migration Apoptosis |
| 基金项目:陕西省自然科学基础研究计划项目（2014JM4141）；湖南省自然科学基金项目（14JJ2092） |
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| 中文摘要: |
| 摘要 目的：探讨miR-221对甲状腺乳头癌生物学特性的影响。方法：培养人甲状腺乳头癌细胞株BCPAP、K1、TPC-1和正常甲状腺细胞株Nthy-ori 3-1。将实验分为四组：A：miR-221模拟物组；B组：miR-221抑制物组；C：无关序列组；D:空白对照组。RT-qPCR的方法检测miR-221在各个细胞中的表达以及转染后各组细胞的表达；MTT实验检测转染后各组细胞的增殖；划痕实验检测转染后各组细胞的迁移能力；流式细胞仪检测转染后各组细胞的凋亡情况。结果：RT-qPCR检测miR-221在三个细胞株的表达情况显示，miR-221甲状腺乳头癌细胞株TPC-1的表达最高，因此选择TPC-1作为后续的研究；miR-221在转染后各组细胞的表达量显示，转染miR221模拟物的miR221的表达显著高于空白对照组，转染miR221抑制物的miR221的表达显著低于空白对照组（P< 0.001）；MTT实验结果显示，转染miR-221模拟物组细胞的增殖速度最快，转染miR-221抑制物组细胞的增殖速度最慢，miR-221模拟物组和miR-221抑制物组细胞从第三天开始与空白对照组有显著差异（P< 0.01），无关对照组与空白对照组无显著差异(P> 0.05)；划痕实验结果显示，转染miR-221模拟物组细胞的迁移数显著高于空白对照组，转染miR-221抑制物组细胞的迁移数显著低于空白对照组（P< 0.01)，无关对照组与空白对照组无显著差异(P> 0.05)；流式细胞仪结果显示，转染miR-221模拟物组细胞凋亡率显著低于空白对照组(P< 0.01)，转染miR-221抑制组细胞凋亡率显著高于空白对照组（P< 0.001），转染无关对照对细胞凋亡无影响(P> 0.05)。结论：过表达miR-221可促进细胞增殖、迁移，抑制细胞凋亡。抑制miR-221表达可降低细胞增殖、迁移，增加细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of miR-221 on the biological characteristics of papillary thyroid carcinoma. Methods: We cultured human papillary thyroid carcinoma cell lines BCPAP, K1, TPC-1 and normal thyroid cell lines 3-1 Nthy-ori. Four groups were designed in this experiment, A: miR-221 mimics group; B: miR-221 inhibitor group; C: unrelated sequence group; D: blank control group. RT-qPCR method was used to detect the expression of miR-221 in individual cells and expression in transfected cells. The proliferation of cells in each group was determined by MTT assay after transfection. The cells migration ability after transfection in each group of cells was detected by scratch assay. And the flow cytometry was used to detect cells apoptosis after the transfection in each group. Results: The expression of miR-221 was detected by RT-qPCR in three cell lines. The expression of TPC-1 was the highest in the papillary thyroid carcinoma cell line, so the TPC-1 was selected for the follow-up study. After transfection, the expression of miR-221 in mimics group was significantly higher than that of the blank control group, while the expression of miR-221 in miR-221 inhibitor group was significantly lower than that of the blank control group (P < 0.001). MTT assay showed that the proliferation of cells was the fastest in miR-221 mimics group, and the slowest in miR-221 inhibitor group. The cell proliferation speed in the miR-221 mimics group and miR-221 inhibitor group had significant difference from the third day as compared to the blank control group (P < 0.01), but it had no significant difference between the unrelated sequence group and the blank control group. The scratch test results showed that the number of transference cells in miR-221 mimics group were significantly more than in blank control group, and that in miR-221 inhibitor group were much less than in blank control group (P<0.01). No such difference were found between the unrelated control group and blank control group (P>0.05). The flow cytometry results displayed that the apoptosis rate was significantly lower in miR-221 mimics groups than in blank control group, but significantly higher in miR-221 inhibition group than in blank control group (P<0.001). There was no significant difference in cell apoptosis between the blank control group and unrelated sequence group. Conclusion: Over expression of miR-221 can promote cell proliferation and migration, and reduce cell apoptosis. Inhibition of miR-221 expression could reduce the cell proliferation and migration and increase apoptosis. |
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