熊果酸对人喉鳞状细胞癌Hep-2细胞增殖和凋亡的影响及其机制研究

摘要

Objective: To investigate tHe effects and mecHanism of ursoLic acid on tHe proLiferation and apoptosis of LaryngeaL squamous Hep-2 ceLLs. MetHods: THe LaryngeaL squamous Hep-2 ceLLs in LogaritHmic growtH pHase were divided into five groups: tHe bLank controL group, UA groups (12. 5, 25, 50 μmoL/ L), and CispLatin group (6 μmoL/ L).Twenty- four Hours after tHe intervention, tHe ceLLuLar growtH inHibition rate was caLcuLated by MTT, ceLL cycLe and apoptosis rate were anaLyzed by fLow cytometry, tHe expression of Bax mRNA and bcL-2 mRNA were detected by RT-PCR to caLcuLate tHe ratio of Bax/ bcL- 2, and tHe expression of caspase- 3 protein was detected by western bLotting metHod. ResuLts: Compared to tHe bLank controL group, tHe ceLLuLar growtH inHibition rate of UA groups (25, 50 μmoL/L) and CispLatin group (6 μmoL/ L) were significantLy increased, witH increased G0/ G1 and decreased G2/ M, tHe apoptosis rate was significantLy increased; tHe expression of bcL-2 mRNA was significantLy decreased, wHiLe tHe expression of Bax mRNA was significantLy increased , and tHe ratio of Bax/ bcL-2 were significantLy increased; tHe expression of caspase-3 protein were significantLy eLevated; aLL of tHe differences above were significant(P ＜0. 05, P ＜ 0. 01).ConcLusion: UrsoLic Acid Has inHibitive effects on tHe proLiferation and acceLerating effects on tHe apoptosis of LaryngeaL squamous Hep-2 ceLLs; wHicH is perHaps reLated to its effects of aLtering ceLL cycLe, aLtering tHe expression of apoptosisreLated genes and proteins.%目的:研究熊果酸(Ursolic Acid, UA)对人喉鳞状细胞癌Hep-2细胞增殖和凋亡的影响及其机制.方法:体外培养并取对数生长期的人喉鳞状细胞癌Hep-2细胞分为空白对照组、UA(12.5、25、50 μmol/L)组和顺铂(6 μmol/L)组,每组设10个复孔.药物干预24h后,采用MTT比色法测定增值抑制率,通过流式细胞术检测细胞周期、观察细胞凋亡状况并计算凋亡率,逆转录-PCR(RT-PCR)法检测bcl-2 mRNA和Bax mRNA表达并计算Bax/bcl-2表达比值,Western blotting法检测caspase-3蛋白表达.结果:UA(25、50 μg/mL)组和顺铂组Hep-2细胞增殖抑制率较空白对照组显著升高,G0/G1期显著增长、G2/M期显著缩短,细胞凋亡率显著升高,细胞中bcl-2 mRNA表达显著下调且Bax mRNA显著上调、Bax/bcl-2表达比值显著升高,caspase-3蛋白表达显著上调,上述差异均具有统计学意义(P<0.05,P<0.01).结论:UA具有抑制人喉鳞状细胞癌Hep-2细胞增殖并促进其凋亡的作用,其机制可能与UA能够改变细胞周期以及调节凋亡相关基因和蛋白表达有关.