摘要
目的探讨结肠癌SW620细胞株中微小核糖核酸-200c(miR-200c)的表达对5-氟尿嘧啶(5-FU)化学敏感性的影响。方法实验分为miR-200c类似物转染SW620细胞株组(A组)、miR-200c类似物阴性对照转染组(B组)、miR-200c抑制物转染SW620细胞株组(C组)、miR-200c抑制物阴性对照转染组(D组)和空白对照组(E组)。采用RT-PCR、MTS及流式细胞术分别检测转染后细胞中miR-200c的表达、5-FU半数抑制浓度(IC50)及细胞凋亡;Western blot检测转染后细胞中磷酸化丝氨酸/苏氨酸激酶(p-Akt)的表达。结果与B组相比,A组转染24h后miR-200c的表达水平提高了(35.94±0.39)倍(P<0.05),5-FU的IC50提高[(1512.67±0.48)μg/ml vs.(926±0.70)μg/ml](P<0.05),5-FU促细胞凋亡的作用降低[(5.01±0.25)%vs.(8.07±0.16)%](P<0.05)。与D组相比,C组miR-200c的表达水平降低了(19.61±0.86)倍(P<0.05),5-FU的IC50降低[(740±0.60)μg/ml vs.(941±0.59)μg/ml](P<0.05),5-FU促细胞凋亡作用提高[(12.23±0.60)%vs.(8.22±0.18)%](P<0.05)。转染48h后,A组p-Akt的表达较B组升高(P<0.05),C组p-Akt表达较D组降低(P<0.05)。结论在结肠癌SW620细胞中,降低miR-200c的表达可提高结肠癌细胞对5-FU的化学敏感性,可能与miR-200c调控Akt的磷酸化有关。
Objective To explore the effects of microRNA 200c （miR-200c） expression on chemosensitivity of SW620 colon cancer cells to 5-FU. Methods Colon cancer cell line SW620 cells were transfected with mimics（group A）, miR-200c mimics negative control （group B）, miR-200c inhibitor（group C）,miR-200c inhibitor negative control（group D） and blank control（group E）. The expression of miR-200c, half inhibition concentration（IC50 ） of 5-FU, the apoptosis rate and that of p-Akt protein expression were detected by RT-PCR, MTS assay, flow cytometry and Western blot, respectively. Results Compared to group B after transfection for 24 hours,the miR-200c expression of group A was increased by （35.94±0. 39） times（P〈0. 05） ,the IC50 of 5-FU was higher[（1512.67±0.48）μg/ml vs. （926±0.70） μg/ml] （P〈0. 05） ,the apoptosis rate was lower[（5.01±0. 25） % vs. （8. 07±0. 16）% ] （P〈0. 05）. Compared to group D, the miR-200c expression of group A was decreased by （19.61±0. 86） times （P〈0. 05）,the ICs0 of 5-FU was lower[-（740 ±0. 60） μg/ml vs. （941±0.59） μg/ml] （P〈0. 05）, the apoptosis rate was higher[（12.23 ± 0.60）% vs. （8.22 ± 0. 18）%]（P〈0. 05）. After transfection for 24 hours, the expression of p-Akt in group A was higher than that in group B （P〈0. 05）, which in group C was lower than that in group D （P〈0. 05）. Conclusion The chemosensitivity of SW620 colon cancer cells to 5-FU can be elevated by reducing the expression of miR-200c,which may be related to the regulation and control of Akt phosphorylation.
出处
《江苏医药》
CAS
北大核心
2014年第11期1245-1248,共4页
Jiangsu Medical Journal
基金
江苏省自然科学基金(BK2010336)
