CAJ | 학술논문

目的观察曲古菌素A(TSA)对肾癌细胞7860增殖抑制作用及对组蛋白去乙酰化酶1(HDAC1)、基质金属蛋白酶9(MMP9)表达的影响.方法噻唑蓝(MTT)比色法测定不同浓度(100、200、400、800nmol/L)、不同时间(12、24、36、48 h)TSA作用后7860细胞的抑制率.吖啶橙/溴乙锭(AO/EB)双重染色检测细胞凋亡.逆转录-聚合酶链反应(RT-PCR)及Western blot检测HDAC1、MMP9的表达.结果不同浓度TSA作用24h后,细胞生长抑制率分别为32%、45%、58%、62%.TSA作用后部分细胞发生凋亡形态改变.HDAC1、MMP9基因在TSA作用后mRNA表达降低(P<0.01).与空白组、阴性组比较,HDAC1蛋白在TSA作用后表达分别下调18.6%～87.5%(P<0.01)、21.3%～91.4%(P<0.01),MMP9蛋白表达分别下调24.6%～95.7%(P<0.01)、20.8%～89.2%(P<0.01).结论 TSA可抑制肾癌细胞增殖,并可能通过下调HDAC1、MMP9的表达抑制细胞的增殖.
목적 관찰곡고균소A(TSA)대신암세포7860증식억제작용급대조단백거을선화매1(HDAC1)、기질금속단백매9(MMP9)표체적영향.방법 새서람(MTT)비색법측정불동농도(100、200、400、800nmol/L)、불동시간(12、24、36、48 h)TSA작용후7860세포적억제솔.아정등/추을정(AO/EB)쌍중염색검측세포조망.역전록-취합매련반응(RT-PCR)급Western blot검측HDAC1、MMP9적표체.결과 불동농도TSA작용24h후,세포생장억제솔분별위32%、45%、58%、62%.TSA작용후부분세포발생조망형태개변.HDAC1、MMP9기인재TSA작용후mRNA표체강저(P<0.01).여공백조、음성조비교,HDAC1단백재TSA작용후표체분별하조18.6%～87.5%(P<0.01)、21.3%～91.4%(P<0.01),MMP9단백표체분별하조24.6%～95.7%(P<0.01)、20.8%～89.2%(P<0.01).결론 TSA가억제신암세포증식,병가능통과하조HDAC1、MMP9적표체억제세포적증식.
Objective To study the depressant effect on proliferation and the expression of histone deacetylase 1 (HDAC1) and matrix metalloproteinase 9 (MMP9) when the trichostatin A (TSA) was added to human renal carcinoma cell line 7860. Methods Four concentrations of TSA (100, 200, 400, 800 nmol/L) on7860 at different time durations of 12, 24, 36, 48 h were applied. The proliferation of 7860 was determined by methyl thiazolyl tetrazolium (MTT) method. The apoptosis was observed by AO/EB staining. The expression of HDAC1 and MMP9 in 7860 was examined by reverse transcription-polymerase chain reaction (RT-PCR) with TSA (400 nmol/L) at 24 h. Under the same testing condition, the protein expression level of HDAC1 and MMP9 in 7860 was detected by Western blotting, and the data were analyzed. Results The inhibition ratio was 32% , 45% , 58% , 62% in 7860 cells treated with TSA of 100, 200, 400, 800 nmol/L at 24 h. The apoptosis rate was increased after TSA treatment. The expression of HDAC1 and MMP9 mRNA in cancer cells was reduced after TSA treatment. The expression of HDAC1 protein was reduced by 18. 6%-87. 5% (P<0.01) and 21. 3%-91.4% (P<0.01), and that of MMP9 protein was decreased by 24. 6% -95. 7% (P<0.01) and 20. 8% -89. 2% (P<0.01) respectively as compared with control group and negative group. Conclusion TSA could inhibit the expression of HDAC1 and MMP9, and proliferation of 7860 simultaneously.