南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
7期
1000-1004
,共5页
毛楠%何关生%饶进军%吕琳
毛楠%何關生%饒進軍%呂琳
모남%하관생%요진군%려림
肺肿瘤%顺铂%耐药性%Bmi-1%衰老%细胞周期
肺腫瘤%順鉑%耐藥性%Bmi-1%衰老%細胞週期
폐종류%순박%내약성%Bmi-1%쇠로%세포주기
lung neoplasm%cisplatin%drug-resistance%Bmi-1%cell senescence%cell cycle
目的：探讨沉默Bmi-1表达在逆转人肺癌顺铂耐药性的作用及机制。方法以siRNA沉默Bmi-1表达，MTT法检测细胞顺铂敏感性，流式细胞术检测细胞周期分布与凋亡，β-半乳糖酐酶染色法检测细胞的衰老，Western blot法检测Bmi-1、P14ARF、P16INK4a、P53、P21、Rb、ubi-H2AK119蛋白表达。结果肺癌顺铂耐药株A549/DDP细胞的Bmi-1表达明显高于A549细胞；沉默Bmi-1的表达显著增加耐药株对顺铂的敏感性（IC50从40.3±4.1μmol/L降到18.3±2.8μmol/L，P<0.01），细胞G0/G1期比值显著升高[从（48.9±2.3）%升到（78.7±7.6）%，P<0.01]；细胞β-半乳糖酐酶染色阳性，呈衰老特征。细胞衰老通路的P14ARF、P16INK4a、P53、P21、Rb表达上调，ubi-H2AK119表达下调。结论沉默Bmi-1表达能诱导A549/DDP细胞衰老，逆转对顺铂的耐药性，其作用机制可能与降低ubi-H2AK119表达、调节INK4a/ARF/Rb衰老通路有关。
目的：探討沉默Bmi-1錶達在逆轉人肺癌順鉑耐藥性的作用及機製。方法以siRNA沉默Bmi-1錶達，MTT法檢測細胞順鉑敏感性，流式細胞術檢測細胞週期分佈與凋亡，β-半乳糖酐酶染色法檢測細胞的衰老，Western blot法檢測Bmi-1、P14ARF、P16INK4a、P53、P21、Rb、ubi-H2AK119蛋白錶達。結果肺癌順鉑耐藥株A549/DDP細胞的Bmi-1錶達明顯高于A549細胞；沉默Bmi-1的錶達顯著增加耐藥株對順鉑的敏感性（IC50從40.3±4.1μmol/L降到18.3±2.8μmol/L，P<0.01），細胞G0/G1期比值顯著升高[從（48.9±2.3）%升到（78.7±7.6）%，P<0.01]；細胞β-半乳糖酐酶染色暘性，呈衰老特徵。細胞衰老通路的P14ARF、P16INK4a、P53、P21、Rb錶達上調，ubi-H2AK119錶達下調。結論沉默Bmi-1錶達能誘導A549/DDP細胞衰老，逆轉對順鉑的耐藥性，其作用機製可能與降低ubi-H2AK119錶達、調節INK4a/ARF/Rb衰老通路有關。
목적：탐토침묵Bmi-1표체재역전인폐암순박내약성적작용급궤제。방법이siRNA침묵Bmi-1표체，MTT법검측세포순박민감성，류식세포술검측세포주기분포여조망，β-반유당항매염색법검측세포적쇠로，Western blot법검측Bmi-1、P14ARF、P16INK4a、P53、P21、Rb、ubi-H2AK119단백표체。결과폐암순박내약주A549/DDP세포적Bmi-1표체명현고우A549세포；침묵Bmi-1적표체현저증가내약주대순박적민감성（IC50종40.3±4.1μmol/L강도18.3±2.8μmol/L，P<0.01），세포G0/G1기비치현저승고[종（48.9±2.3）%승도（78.7±7.6）%，P<0.01]；세포β-반유당항매염색양성，정쇠로특정。세포쇠로통로적P14ARF、P16INK4a、P53、P21、Rb표체상조，ubi-H2AK119표체하조。결론침묵Bmi-1표체능유도A549/DDP세포쇠로，역전대순박적내약성，기작용궤제가능여강저ubi-H2AK119표체、조절INK4a/ARF/Rb쇠로통로유관。
Objective To investigate the effect of silencing Bmi-1 expression in reversing cisplatin resistance in human lung cancer cells and explore the possible mechanisms. Methods Cisplatin-resistant A549/DDP cells with small interference RNA (siRNA)-mediated Bmi-1 expression silencing were examined for cisplatin sensitivity using MTT assay and alterations in cell cycle distribution and apoptosis with flow cytometry, and the changes in cell senescence was assessed using β-galactosidase staining. The protein expressions of Bmi-1, P14ARF, P16INK4a, P53, P21, Rb and ubi-H2AK119 in the cells were determined with Western blotting. Results A549/DDP cells showed significantly higher Bmi-1 expression than A549 cells. After siRNA-mediated Bmi-1 silencing, A549/DDP cells showed significantly enhanced cisplatin sensitivity with an increased IC50 from 40.3± 4.1μmol/L to 18.3 ± 2.8μmol/L (P<0.01) and increased cell percentage in G0/G1 phase from (48.9 ± 2.3)%to (78.7 ± 7.6)%(P<0.01). Silencing Bmi-1 did not cause significant changes in the cell apoptosis rate but induced obvious senescence phenotype in A549/DDP cells with down-regulated expression of ubi-H2AK119 and up-regulated expressions of P14ARF, P16INK4a, P53,P21 and Rb. Conclusion Silencing Bmi-1 by RNA interference can induce cell senescence and resensitize A549/DDP cells to cisplatin possibly by regulating INK4a/ARF/Rb senescence pathway.
