Tetrandrine enhances radiosensitivity through the CDC25C/CDK1/cyclin B1 pathway in nasopharyngeal carcinoma cells

The increasing resistance of nasopharyngeal carcinoma to irradiation makes the exploration of effective radiosensitizers necessary. Tetrandrine is known to be an antitumor drug, but little is known regarding its radiosensitization effect on nasopharyngeal carcinoma. We investigated the effect of combined treatment of irradiation and maximum non-cytotoxic doses of tetrandrine on the nasopharyngeal carcinoma cell lines CNE1 and CNE2. The maximum non-cytotoxic doses of tetrandrine in CNE1 and CNE2 cells were assessed using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The radiosensitization of cells receiving the maximum non-cytotoxic doses of tetrandrine was assessed by evaluating cell proliferation and DNA damage repair using MTT, clonogenic, comet assays and detection of caspase-3 and phosphorylated histone H2AX (γ-H2AX). The cell cycle was assessed by flow cytometry, and protein expression was detected by western blot analysis. The maximum non-cytotoxic doses of tetrandrine in CNE1 and CNE2 cells were 1.5 μmol/L and 1.8 μmol/L, respectively. When cells were exposed to irradiation and the maximum non-cytotoxic doses of tetrandrine, the survival fraction was decreased. DNA damage and γ-H2AX levels markedly increased. Moreover, tetrandrine abrogated the G2/M phase arrest caused by irradiation. Combined treatment with the maximum non-cytotoxic dose of tetrandrine and irradiation caused suppression of the phosphorylation of CDK1 and CDC25C and increase in the expression of cyclin B1. The study in vivo also showed that the maximum non-cytotoxic dose of tetrandrine could reduce tumor growth in xenograft tumor model. Our results suggest that the maximum non-cytotoxic dose of tetrandrine can enhance the radiosensitivity of CNE1 and CNE2 cells and that the underlying mechanism could be associated with abrogation of radiation-induced G2/M arrest via activation of the CDC25C/CDK1/Cyclin B1 pathway.     

三氧化二砷诱导CNE1凋亡及其对细胞周期的影响

目的 研究三氧化二砷对人鼻咽癌CNE1细胞凋亡及其细胞周期的影响。方法 应用形态学观察、原位末端标记法(TUNEL)、流式细胞术等方法对三氧化二砷诱导的鼻咽癌细胞CNE1进行检测和观察。结果 一定浓度三氧化二砷能诱导CNE1细胞凋亡,凋亡细胞具有典型的凋亡形态特征,TUNEL原位检测有典型凋亡细胞,流式细胞仪检测有凋亡峰,G2／M期比例升高,呈一定的剂量效应关系。结论 三氧化二砷能诱导人鼻咽癌CNE1细胞株凋亡及阻止细胞周期进展的作用。     

TRAIL联合多西紫杉醇对人鼻咽癌CNE2细胞的体外作用研究

目的:研究肿瘤坏死因子相关凋亡配体(TRAIL)联合化疗药物多西紫杉醇对鼻咽癌CNE2细胞凋亡诱导作用。方法:应用MTT法检测不同浓度多西紫杉醇的抗癌活性,计算其亚毒性剂量,流式细胞仪检测多西紫杉醇及TRAIL单独或者联合作用于鼻咽癌CNE2细胞后的细胞凋亡发生率,TUNEL法观察细胞凋亡发生情况。结果:鼻咽癌细胞对TRAIL的作用敏感,多西紫杉醇可以增强其凋亡诱导作用。结论:TRAIL与多西紫杉醇具有协同抗鼻咽癌作用,有望应用于鼻咽癌的临床治疗。     

Suppression of bcl-2 gene by RNA interference increases chemosensitivity to cisplatin in nasopharyngeal carcinoma cell line CNE1

To explore the effect of suppressing BCL-2 expression using RNA interference (RNAi) technique in nasopharyngeal carcinoma cell line CNE1. CNE1 cell lines stably expressing shRNAs targeted bcl-2 and GL3 gene were established and gene expression inhibition was assessed by Western blotting analysis. The effect of suppressing bcl-2 by RNAi on cell growth was studied, the apoptosis induction and the sensitization of CNE1 cells to cisplatin were quantified by MTT assay and flow cytometry. The results showed that: stable transfection of CNE1     

JNK相互作用蛋白通过JNK途径影响鼻咽癌的增殖和凋亡

EB病毒编码的瘤蛋白潜伏膜蛋白（LMP1）所介导的活化蛋白（AP-1）信号转导途径在细胞增殖、分化、转化与凋亡方面发挥着重要作用.越来越多的证据表明,AP-1信号转导通路中上游激酶JNK在鼻咽癌的发生发展过程中起着重要作用.最近克隆出来的JNK相互作用蛋白(JIP-1)是一种能抑制JNK核移位的胞浆锚蛋白.为探讨JIP在LMP1调控AP-1信号通路中的作用机制,采用间接免疫荧光法和报告基因法,发现JIP通过有效地抑制磷酸化的JNK从胞浆移位入核,从而抑制LMP1上调的AP-1活性.同时,JIP导入鼻咽癌细胞中,MTT法发现JIP能够明显抑制鼻咽癌细胞的生长.进一步发现转染JIP后细胞的集落形成率与对照组相比大约降低了53.6%,也抑制了细胞. 提示JIP可明显抑制细胞的增殖作用.进一步采用流式细胞术分析,结果发现JIP引起细胞G1/S期细胞阻滞,说明JIP是抑制细胞增殖的重要调节子.进一步采用流式细胞术定量发现,转染JIP后细胞的24 h凋亡百分率由1.25%上升到8.25%,上升约6.6倍,48 h由1.04%上升到31.45%,上升约30倍. 采用激光共聚焦显微镜发现,转染JIP后细胞核发生显著变化,核质由均匀状态固缩成高凝集状态,形成了典型的胞膜体.提示JIP可有效地促进细胞凋亡.结果表明,JIP可通过抑制活化的JNK核移位,降低LMP1所介导的AP-1信号通路.并进一步发现JIP可有效地抑制细胞增殖和细胞凋亡,从而提示JIP可作为新的治疗肿瘤潜在靶分子.     

槐米中天然产物槲皮素抗人鼻咽癌CNE1细胞活性研究

本研究以槐米为原材料提纯天然产物槲皮素,以人鼻咽癌细胞系CNE1为试验对象,探索天然产物槲皮素对人鼻咽癌细胞的增殖抑制效应及凋亡诱导效应。研究方法采用超声醇提法从槐米中提取芸香苷,再经酸水解和重结晶制备槲皮素精制品;采用槲皮素标准曲线比色法检测槲皮素精制品中槲皮素的纯度;采用MTT比色法进行细胞毒性试验,检测槲皮素精制品对人鼻咽癌细胞CNE1增殖的抑制效应及半数抑制浓度(IC50);采用流式细胞术结合AnnexinV、PI双染色法进行细胞凋亡试验,检测槲皮素精制品对CNE1细胞凋亡的诱导效应。结果表明,从槐米中提纯的天然产物槲皮素精制品,能呈剂量依赖性地抑制人鼻咽癌细胞CNE1增殖并诱导其凋亡。综上所述,应用超声醇提法,从槐米中制备的天然产物槲皮素精制品,不仅纯度高,且有着较优的抗癌活性,进一步明确了槐米等中药材开发天然产物有效成分,服务于人类健康的潜在应用价值和社会与经济效益。     

参杞合剂对人鼻咽癌细胞CNE细胞周期及凋亡的影响

目的研究参杞合剂(SQ)在体外对CNE细胞周期及凋亡的影响。方法应用MTT法观察参杞合剂对细胞的抑制作用,流式细胞术观察不同浓度参杞合剂作用不同时间后CNE细胞周期的改变,电镜结合DNA电泳分析参杞合剂诱导凋亡的作用。结果SQ对CNE细胞生长有明显抑制作用,且其作用强度呈现出对浓度和时间的依赖性。CNE细胞在SQ作用下随着时间的延长和浓度的增加,G0/G1期比率下降,S期比率升高,出现S期阻滞。0.0625 g.生药/ml的SQ作用48 h后诱导出凋亡,凋亡率随着浓度的增加、时间的延长而增加,电镜下可见典型凋亡小体。琼脂糖凝胶电泳呈现出凋亡特征性的DNA条带。结论参杞合剂可直接杀伤肿瘤细胞,其机制可能通过阻滞细胞周期S期,诱导肿瘤细胞凋亡实现的。     

鼻咽癌相关新基因NPCEDRG表达对CNE2细胞生长的影响

为研究鼻咽癌相关新基因NPCEDRG的功能,探讨其对鼻咽癌细胞生长特性的影响,利用Tet-on调控系统,建立受强力霉素(deoxycycline,Dox)诱导NPCEDRG基因表达的CNE2细胞系.运用RT-PCR选择背景表达低、诱导活性高的细胞克隆,以不同浓度Dox诱导CNE2/Tet/TRE-NPCEDRG细胞,确定Dox的最佳诱导浓度.借助形态学观察、细胞生长曲线、软琼脂克隆形成试验和流式细胞仪分析等方法,对Dox诱导NPCEDRG高表达后CNE2细胞的生物学行为进行了检测.结果显示,NPCEDRG高表达后CNE2细胞增殖速度显著减慢(P<0.05),克隆形成能力显著降低(P<0.01),瘤细胞群体中处于G0/G1期细胞数增加,S期细胞数减少,细胞阻滞于G0/G1期.Tet调控NPCEDRG基因表达CNE2细胞系成功建立,恢复NPCEDRG表达能部分逆转CNE2的恶性表型,证明NPCEDRG是一个鼻咽癌相关的抑瘤基因.     

苦参碱对鼻咽癌细胞凋亡及凋亡相关基因p53的表达研究

目的：探讨苦参碱对体外培养的人鼻咽癌细胞增殖、凋亡及凋亡相关基因p53 mRNA和蛋白表达的影响,初步探讨苦参碱诱导人鼻咽癌细胞凋亡的可能机制。方法：采用MTT法检测不同浓度苦参碱（0、0.25、0.5、1、1.5、2 mg/ml）对CNE1、CNE2细胞增殖的影响;采用荧光定量PCR法检测这些浓度的苦参碱处理48 h后CNE2细胞p53 mRNA的变化;Western Blot检测其蛋白的变化情况。结果：MTT结果显示苦参碱具有抑制CNE1、CNE2细胞体外增殖作用,其抑制率存在浓度、时间依赖性。荧光定量PCR及Western Blot检测结果显示,苦参碱抑制CNE2细胞p53 mRNA和蛋白的表达,且亦呈浓度依赖性。结论：苦参碱抑制CNE2细胞的增殖,诱导细胞凋亡,呈现浓度、时间依赖性,其作用与抑制CNE2细胞中p53基因和蛋白的表达密切相关。     

Deoxyelephantopin from Elephantopus scaber L. induces cell-cycle arrest and apoptosis in the human nasopharyngeal cancer CNE cells

Deoxyelephantopin (ESD), a naturally occurring sesquiterpene lactone present in the Chinese medicinal herb, Elephantopus scaber L. exerted anticancer effects on various cultured cancer cells. However, the cellular mechanisms by which it controls the development of the cancer cells are unavailable, particularly the human nasopharyngeal cancer CNE cells. In this study, we found that ESD inhibited the CNE cell proliferation. Cell cycle arrest in S and G2/M phases was also found. Western blotting analysis showed that modulation of cell cycle regulatory proteins was responsible for the ESD-induced cell cycle arrest. Besides, ESD also triggered apoptosis in CNE cells. Dysfunction in mitochondria was found to be associated with the ESD-induced apoptosis as evidenced by the loss of mitochondrial membrane potential (ΔΨm), the translocation of cytochrome c, and the regulation of Bcl-2 family proteins. Despite the Western blotting analysis showed that both intrinsic and extrinsic apoptotic pathways (cleavage of caspases-3, -7, -8, -9, and -10) were triggered in the ESD-induced apoptosis, additional analysis also showed that the induction of apoptosis could be achieved by the caspase-independent manner. Besides, Akt, ERK and JNK pathways were found to involve in ESD-induced cell death. Overall, our findings provided the first evidence that ESD induced cell cycle arrest, and apoptosis in CNE cells. ESD could be a potential chemotherapeutic agent in the treatment of nasopharyngeal cancer (NPC).     

TNFalpha-mediated cell death is independent of cdc25A

Tumor necrosis factor (TNFs) have been shown to be synthesized by ovarian carcinomas, and may therefore affect tumor cells in an autocrine manner. Therefore, we investigated the effects of recombinant TNFs on ovarian carcinoma cells N.1 and examined expression of the proto-oncogenes c-myc and cdc25A which are known to play a prominent role in apoptosis. TNFalpha elicited apoptosis in N.1 cells within 72 h which was shown by typical morphological changes, DNA fragmentation and signature type cleavage of poly(ADP-ribose) polymerase into a 89 kDa proteolytic peptide. TNFalpha-induced apoptosis was accompanied by constitutive c-Myc expression, although the mRNA level of phosphatase cdc25A was suppressed within 24 h of TNFalpha treatment and the protein level decreased after 48 h. Cdc25A tyrosine phosphatase is an activator of the cdk2-cyclin E complex which allows for cell cycle progression. As expected, we found TNFalpha-mediated Cdc25A down-regulation to inhibit Cdk2 activity. Cdc25A suppression was related to TNFalpha-induced apoptosis but not to a TNFalpha-induced G0 arrest because cyclin D1 expression was unaffected and the gene gas6 (growth arrest specific 6) was not induced. Arresting cells by treatment with genistein prevented TNFalpha-triggered apoptosis and inhibited c-myc expression. TNFalpha-induced apoptosis is not accompanied by cell cycle arrest which may be due to constitutive c-Myc expression, although Cdc25A and Cdk2 activity is also down-regulated. High c-Myc and low Cdc25A activity might present conflicting signals to the cell cycle machinery which are incompatible with cell survival.     

双氢青蒿素调控巨噬细胞增殖和迁移的作用研究

目的:探讨双氢青蒿素在体外对小鼠单核巨噬细胞RAW264.7的增殖、克隆形成、周期、凋亡和迁移的影响。方法:采用梯度浓度(2.5μg/m L, 5μg/m L, 10μg/m L, 20μg/m L)的双氢青蒿素处理RAW264.7细胞,利用CCK8实验检测双氢青蒿素对巨噬细胞增殖能力的影响,利用克隆形成实验检测双氢青蒿素对RAW264.7细胞克隆形成能力的影响,利用流式细胞术检测双氢青蒿素对RAW264.7细胞周期和凋亡的影响,利用划痕修复实验检测RAW264.7细胞迁移能力。结果:CCK8实验结果显示,双氢青蒿素可以显著抑制RAW264.7巨噬细胞的增殖能力,且抑制效果与双氢青蒿素的浓度呈正相关性。克隆形成实验结果显示,双氢青蒿素可以抑制细胞的克隆形成能力。双氢青蒿素处理使RAW264.7细胞G0/G1期比例显著升高,S期与G2/M期细胞比例显著降低。双氢青蒿素对巨噬细胞凋亡具有诱导作用,且凋亡诱导作用呈现浓度依赖的特性。划痕修复实验结果显示,双氢青蒿素可以显著抑制RAW264.7巨噬细胞的迁移能力。结论:双氢青蒿素可以导致巨噬细胞的细胞周期G0/G1阻滞,并且诱导细胞凋亡,对巨噬细胞增殖和迁移具有抑制作用。     

PXD101对口腔鳞癌HN-6细胞增殖及凋亡的初步研究

目的:本实验探讨组蛋白去乙酰化酶抑制剂PXD101对口腔鳞癌HN-6细胞的增殖、细胞凋亡及细胞周期的影响。方法:PXD101对口腔鳞癌HN-6细胞进行干预,倒置相差显微镜观察细胞形态学改变;MTT法检测PXD101对HN-6细胞的增殖影响;Annexin-V-FITC/PI双染流式细胞仪定量检测细胞凋亡;流式细胞仪分析细胞周期。结果:PXD101可明显抑制HN-6细胞的生长(P0.05),呈时间剂量依赖性。倒置相差显微镜下观察对照组细胞贴壁,形态呈多边形,生长活跃;实验组细胞脱壁,变小,细胞核皱缩。绘制细胞生长曲线示,随着PXD101的浓度和作用时间的增加,HN-6细胞生长明显受到抑制,各实验组细胞生长抑制率与对照组比较,P0.05差别有统计学意义。1μmol/LPXD101作用24 h,48 h细胞总凋亡率分别为20.9%、38.6%,与对照组相比有统计学意义(P0.05);HN-6细胞周期阻滞于G0/G1期,与对照组相比,P0.05差别有统计学意义。结论:PXD101体外实验能显著抑制人口腔舌癌HN-6的细胞增殖,诱导细胞凋亡。     

Effects of 2-methoxyestradiol on proliferation, apoptosis and gene expression of cyclin B1 and c-Myc in esophageal carcinoma EC9706 cells

2-Methoxyestradiol (2-ME) is an endogenous metabolite of 17β-estradiol. In this study, we determined the antitumour activities of 2-ME on the well-differentiated EC9706 esophageal carcinoma cells in vitro. 2-ME had a strong antiproliferative effect on EC9706 cells and caused an increase in the population of apoptotic cells, detected by flow cytometry. A significant number of cells were blocked in the G(2)/M phase of the cell cycle. 2-ME-treated cells demonstrated an increase in cyclin B1 and c-Myc protein levels, as well as an increase in the percentage of G(2)/M phase. Their up-regulation may be involved in 2-ME-induced apoptosis and G(2)/M cell cycle arrest of the EC9706 cells, and it precedes the onset of apoptosis.     

Human immunodeficiency virus type 1 Vpr induces apoptosis following cell cycle arrest.

The human immunodeficiency virus type 1 (HIV-1) vpr gene encodes a protein which induces arrest of cells in the G2 phase of the cell cycle. Here, we demonstrate that following the arrest of cells in G2, Vpr induces apoptosis in human fibroblasts, T cells, and primary peripheral blood lymphocytes. Analysis of various mutations in the vpr gene revealed that the extent of Vpr-induced G2 arrest correlated with the levels of apoptosis. However, the alleviation of Vpr-induced G2 arrest by treatment with the drug pentoxifylline did not abrogate apoptosis. Together these studies indicate that induction of G2 arrest, but not necessarily continued arrest in G2, was required for Vpr-induced apoptosis to occur. Finally, Vpr-induced G2 arrest has previously been correlated with inactivation of the Cdc2 kinase. Some models of apoptosis have demonstrated a requirement for active Cdc2 kinase for apoptosis to occur. Here we show that accumulation of the hypophosphorylated or active form of the Cdc2 kinase is not required for Vpr-induced apoptosis. These studies indicate that Vpr is capable of inducing apoptosis, and we propose that both the initial arrest of cells and subsequent apoptosis may contribute to CD4 cell depletion in HIV-1 disease.     

LATS1过表达对肺癌A549细胞增殖及周期的影响

通过过表达手段上调大肿瘤抑制因子1（1arge tumor suppressor gene 1,LATS1）基因在A549细胞中的表达,研究LATS1对A549细胞生长和细胞周期调控的作用。构建过表达LATS1基因的慢病毒载体,转染A549N胞株,采用RT-PCR和蛋白质印迹法检测转染后A549细胞中LATS1、YAPmRNA和蛋白的表达效率;流式细胞术检测细胞凋亡、周期情况：CCK-8检测细胞的增殖水平变化。结果发现,过表达LATS1慢病毒载体转染A549细胞株后,LATS1mRNA及蛋白表达水平高于未处理组及转染空载体组,YAPmRNA及蛋白表达水平低于未处理组及转染空载体组;过表达LATS1慢病毒转染后,A549细胞增殖率从第五天开始低于对照组（P〈0．05）,过表达组细胞G1期比例明显增高（P〈0．05）,凋亡率明显增加（P〈0．05）,差异均有统计学意义。以上结果提示,LATS1可通过下调YAP的表达水平促进A549细胞的凋亡,诱导G1期阻滞,降低细胞的增殖能力。     

罗汉果醇抗肿瘤活性及其作用机制研究

罗汉果醇是罗汉果皂苷的苷元,有研究报道罗汉果皂苷V具有防癌抑癌作用。该研究采用噻唑蓝实验( MTT法)检测罗汉果醇对不同肿瘤细胞增殖的抑制情况,以及不同浓度的罗汉果醇对CNE1细胞的增殖抑制率;应用细胞克隆形成实验进一步验证罗汉果醇对CNE1细胞增殖的抑制作用;采用Annexin V/PI 双染法检测罗汉果醇对CNE1细胞凋亡的影响;以实时定量PCR技术检测罗汉果醇对CNE1细胞中Caspase-3、Sur-vivin、Bax和Bcl-2基因的mRNA 表达水平的影响。结果表明：罗汉果醇能显著抑制DU145、HepG2、A549、CNE1、CNE2细胞的增殖,其中对CNE1细胞增殖的抑制作用最为显著,并呈剂量依赖性,其半数抑制浓度IC50为(81.48±4.73)μmol·L-1;通过对CNE1细胞进一步的克隆形成实验,也验证了这一点;Annexin V/PI 双染法可见随着浓度的增加,凋亡比例增加;实时定量PCR技术检测显示罗汉果醇处理后,促凋亡基因Caspase-3、Bax的表达增加,抗凋亡基因Survivin、Bcl-2的表达减少。因此,罗汉果醇可能是通过促进Caspase-3、Bax等促凋亡基因和抑制Survivin、Bcl-2等抗凋亡基因的表达,来诱导肿瘤细胞凋亡,进而发挥抗肿瘤活性。     

雷氏大疣蛛毒液对人肝癌HepG2细胞p21基因表达的影响

本文研究了雷氏大疣蛛毒液对人肝癌细胞株HepG2增殖抑制作用及其分子机制。采用XTT法观察到雷氏大疣蛛毒液剂量依赖抑制HepG2细胞增殖;流式细胞仪检测发现,经过雷氏大疣蛛毒液作用的HepG2细胞周期发生明显的选择性改变;RT-PCR方法检测到p21基因表达增强;Western blot检测发现,p21蛋白表达增加。结果提示,雷氏大疣蛛毒液抑制人肝癌细胞HepG2增殖的可能机制之一是使p21基因和蛋白表达增加,G2IM细胞周期被阻滞,从而诱导细胞凋亡。     

Anti-proliferative effects of evodiamine on human prostate cancer cell lines DU145 and PC3

Prostate carcinoma is one of the most common malignant tumors and has become a more common cancer in men. Previous studies demonstrated that evodiamine (EVO) exhibited anti-tumor activities on several cancers, but its effects on androgen-independent prostate cancer are unclear. In the present study, the action mechanisms of EVO on the growth of androgen-independent prostate cancer cells (DU145 and PC3 cells) were explored. EVO dramatically inhibited the growth and elevated cytotoxicity of DU145 and PC3 cells. The flow cytometric analysis of EVO-treated cells indicated a block of G2/M phase and an elevated level of DNA fragmentation. The G2/M arrest was accompanied by elevated Cdc2 kinase activity, an increase in expression of cyclin B1 and phosphorylated Cdc2 (Thr 161), and a decrease in expression of phosphorylated Cdc2 (Tyr 15), Myt-1, and interphase Cdc25C. TUNEL examination showed that EVO-induced apoptosis was observed at 72 h. EVO elevated the activities of caspase 3, 8, and 9 in DU145 cells, while in PC3 cells only the activities of caspase 3 and 9 were elevated. EVO also triggered the processing of caspase 3 and 9 in both DU145 and PC3 cells. We demonstrate that roscovitine treatment result in the reversion of G2/M arrest in response to EVO in both DU145 and PC3. However, inhibitory effect of roscovitine on EVO-induced apoptosis could only be observed in DU145 rather than PC3. In DU145, G2/M arrest might be a signal for initiation of EVO-triggered apoptosis. Whereas EVO-triggered PC3 apoptosis might be independent of G2/M arrest. These results suggested that EVO inhibited the growth of prostate cancer cell lines, DU145 and PC3, through an accumulation at G2/M phase and an induction of apoptosis.     

一种新型PPARr激动剂对人肾癌细胞增殖抑制的实验研究

目的：探讨新型过氧化物酶体增殖激活物受体(PPARr)激动剂DH9 对人肾癌细胞OS-RC-2 的增殖抑制作用。方法：予以不 同浓度的DH9 及罗格列酮作用OS-RC-2 细胞12 h、24 h和48 h,荧光素酶活性检测比较两种药物的PPARr激动效应;MTT 法 检测细胞增殖情况;流式细胞术观察细胞周期;AnnexinV-FITC/PI双染色流式细胞术测定细胞凋亡率;Western blot 检测细胞内 Bax 及Bcl-2等蛋白的变化。结果：不同浓度的DH9 与罗格列酮相比,对PPARr的激动效应DH9明显低于罗格列酮,增殖抑制 作用优于罗格列酮(P＜0.05),并呈现明显的浓度、时间依赖性;加入PPARr抑制剂GW9662 前后DH9 的增殖抑制作用差异无统 计学意义(P＞0.05);DH9 作用细胞48小时后,G0/G1 期细胞比例明显增加(P＜0.05),S期细胞明显减少(P＜0.05)。DH9可诱导细 胞凋亡,伴随Bcl-2 表达的减少以及Bax表达的增加。结论：OS-RC-2 细胞中,DH9 的增殖作用明显优于罗格列酮,且是通过 PPARr非依赖途径实现;DH9 能将OS-RC-2 细胞阻滞在G0/G1 期,并通过影响Bcl-2 和Bax 蛋白表达促进细胞凋亡。