NF-κB在大黄素增强胰腺癌吉西他滨化疗敏感性中的作用

为探讨大黄素增强胰腺癌吉西他滨化疗敏感性的作用及其机制,本研究诱导建立耐100 nmol·L-1吉西他滨的人胰腺癌SW1990细胞株(SW1990/GZ),在体外实验中,应用CCK-8法检测细胞增殖活性;流式细胞术检测细胞凋亡;凝胶电泳迁移率实验(EMSA)检测细胞中NF-κB活性;Western blotting检测细胞中蛋白表达水平;在体内实验中,建立裸鼠胰腺癌原位移植模型;免疫组织化学法检测肿瘤组织中蛋白表达。结果表明,大黄素预处理可显著增强吉西他滨对胰腺癌细胞的生长抑制和诱导凋亡作用;大黄素联合吉西他滨可显著抑制胰腺癌原位移植瘤生长;大黄素还可下调体内外胰腺癌中NF-κB及其调控蛋白Bcl-2和Survivin的表达。提示NF-κB在大黄素增敏胰腺癌吉西他滨化疗中具有重要意义。     

干扰c-myc基因表达增强人胰腺癌细胞对吉西他滨和卡铂的敏感性

目的　探讨干扰c-myc 基因表达对人胰腺癌细胞SW1990 对化疗药物敏感性的影响。方法　采用实时定量PCR和Western blot 检测人胰腺癌细胞系SW1990 中c-myc 基因的干扰效果;MTT 法检测癌细胞对化疗药物吉西他滨和卡铂的敏感性;流式细胞术检测吉西他滨诱导的细胞凋亡情况。结果　c-myc siRNA 显著下调人胰腺癌SW1990 细胞中c-myc 基因的mRNA 和蛋白表达水平。干扰c-myc 表达后,化疗药物吉西他滨和卡铂对SW1990 细胞的半数抑制浓度（IC50）均显著减小（P<0.05）,吉西他滨诱导的SW1990 细胞凋亡率显著升高（P<0.05）。结论　干扰c-myc 表达可增强人胰腺癌细胞株SW1990 对化疗药物吉西他滨和卡铂的敏感性。     

吉西他滨联合卡铂诱导NK/T细胞淋巴瘤细胞株凋亡的研究

目的:研究吉西他滨联合卡铂对NK/T细胞淋巴瘤细胞株(SNK 6)细胞增殖及凋亡的影响。方法:采用细胞增殖-毒性检测法(CCK-8)检测吉西他滨、卡铂、顺铂对SNK 6细胞增殖的影响,算出各自半数抑制浓度(IC 50)值;再用吉西他滨与卡铂或顺铂做联合实验,用金氏公式计算Q值,评价联合用药效应。应用流式细胞仪检测不同药物组对细胞凋亡的影响。蛋白免疫印迹法(Western Blotting)检测各药物组Cleaved-caspase-3、B淋巴细胞瘤-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)等凋亡蛋白的表达水平。结果:吉西他滨、卡铂、顺铂对SNK 6细胞均有较好的抑制作用;联合用药中,吉西他滨+卡铂组与吉西他滨+顺铂组对SNK 6细胞抑制作用相当,两药联合为相加作用。吉西他滨+卡铂组凋亡率稍低于吉西他滨+顺铂组,但差异无统计学意义(P>0.05)。吉西他滨可上调Cleaved-caspase-3蛋白、Bax蛋白的表达水平,下调Bcl-2蛋白的表达水平(P<0.05),吉西他滨与卡铂或顺铂联合时效果更加显著。结论:在体外环境下,吉西他滨联合卡铂可抑制SNK 6细胞增殖并诱导其凋亡,且一定浓度的吉西他滨联合卡铂对SNK 6细胞株的凋亡率与吉西他滨联合顺铂相似。     

吉西他滨对人胰腺癌细胞系BxPC-3的放射增敏作用

目的 探讨吉西他滨对人胰腺癌细胞系BxPC-3的细胞毒性、放射增敏作用.方法 用MTT法测定吉西他滨对BxPC-3细胞生长的作用,集落形成法观察其对BxPC-3细胞的放射增敏作用,流式细胞术分析吉西他滨作用后细胞周期及凋亡变化,免疫细胞化学染色观察Bcl-2、Bax蛋白表达.结果 占西他滨对BxPC-3细胞具有浓度依赖性抑制作用(IC50-106 μg/ml),选择浓度为IC10(0.04μg/ml),可增加X射线对BxPC-3的杀伤作用,放射增敏比SER为1.15.药物浓度为0.04μg/ml时S期细胞比例增高.吉西他滨作用24 h后,细胞Bax表达增加,而Bcl-2表达下降.结论 吉西他滨对人胰腺癌BxPC-3细胞系有细胞毒性作用和一定的放射增敏作用;其机制可能与吉西他滨引起S期阻滞和调节凋亡相关基因的表达有关.     

尼美舒利联合吉西他滨对人非小细胞肺癌NCI-H1975细胞增殖和凋亡的影响

目的研究尼美舒利联合吉西他滨对人非小细胞肺癌NCI-H1975细胞增殖和凋亡的影响。方法采用MTT法测定尼美舒利联合吉西他滨对NCI-H1975细胞生长的抑制作用。流式细胞仪检测尼美舒利(100μmol/L)组、吉西他滨(20 nmol/L)组和联合用药(尼美舒利100μmol/L+吉西他滨20 nmol/L)组对NCI-H1975细胞周期和凋亡的影响。Western blotting检测各组对Bcl-2、Bax、Cleaved caspase-3蛋白表达的影响。结果尼美舒利对NCI-H1975细胞的生长具有抑制作用,且呈剂量相关性,IC50为(298.76±2.79)μmol/L;吉西他滨的IC50为(78.64±3.14)nmol/L,尼美舒利与吉西他滨联合作用后,能明显增加后者对NCI-H1975细胞增殖的抑制作用,二者有明显的协同作用。尼美舒利联合吉西他滨将细胞阻滞在G0/G1期,而S和G2/M期均有所减少。与单药组相比,联合用药组能显著升高细胞的凋亡率(P0.05)。与单药组相比,联合用药组明显下调Bcl-2蛋白的表达(P0.05),上调Bax和Cleaved caspase-3蛋白的表达(P0.05)。结论尼美舒利联合吉西他滨能明显抑制肿瘤细胞的增殖,诱导细胞凋亡,其效果优于单用吉西他滨组,其机制与影响细胞周期调控,下调Bcl-2蛋白表达、上调Bax和Cleaved caspase-3蛋白表达水平有关。     

姜黄素增加HepG2细胞对顺铂敏感性的作用及机制研究

目的　探讨姜黄素对肝癌细胞株HepG2 对顺铂敏感性的影响及其作用机制。方法　采用不同浓度姜黄素（0、2.5、5、7.5、10 μM）及顺铂（10、20、40、80 μM）单用或联用处理HepG2 细胞株后,通过MTT 检测细胞增殖抑制率,流式细胞术检测细胞凋亡情况,Real-time PCR 技术检测Survivin、Cyto-C、Bcl2、Bax 的基因表达情况;western blot 检测Survivin、Cyto-C、cleaved caspase-3、Bcl-2、Bax 的蛋白表达。结果　姜黄素以浓度依赖性方式抑制HepG2细胞株Survivin 的mRNA 表达和蛋白表达（P<0.05）;姜黄素联合顺铂处理的HepG2 细胞增殖抑制率及细胞凋亡率较单用顺铂处理均明显增加（P<0.05）;姜黄素联合顺铂可显著抑制Survivin 的蛋白表达,降低Bcl-2/Bax 比例,增加Cyto-C、cleaved caspase-3 蛋白表达（P<0.05）。结论　姜黄素可增加HepG2 细胞对顺铂的敏感性,可能与其抑制Survivin 的表达,降低Bcl-2/Bax 比例,增加cleaved caspase-3、Cyto-C 蛋白的表达有关。     

吉西他滨联合大黄素对胰腺癌 SW1990细胞多耐药基因?1、 miRNA?1271及上皮-间充质细胞转化的影响

目的探究吉西他滨联合大黄素对胰腺癌 SW1990细胞生长的抑制作用及对多耐药基因 ?1(MDR?1)、 miRNA?1271和上皮-间充质细胞转化( EMT)的影响。方法将胰腺癌 SW1990细胞分为大黄素组( 40 μmol/L)、吉西他滨( 20 μmol/L)、吉西他滨联合大黄素组及对照组(生理盐水),流式细胞仪检测细胞凋亡, MTT法检测细胞增殖, Transwell小室模型检测胰腺癌细胞侵袭能力,实时荧光定量聚合酶链反应( qRT?PCR)检测 MDR?1、miRNA?1271及 EMT相关标志物( TWIST1、ZEB1、E?cadhcrin) mRNA表达,流式细胞仪检测 MDR?1编码的 P糖蛋白( P?gp)阳性率,蛋白质免疫印迹法检测肿瘤组织凋亡相关蛋白［B淋巴细胞瘤 2(Bcl?2)及其相关 X蛋白( Bax)、胱天蛋白酶 3(Caspase?3)及 Survivin］及 EMT相关标志物蛋白含量。结果联合组可显著抑制胰腺癌 SW1990细胞增殖和侵袭, SW1990细胞凋亡率显著更高,与其他三组比较差异有统计学意义( P＜0.05)。联合组 MDR?1 mRNA显著降低, miRNA?1271及 E?cadhcrin mRNA显著升高,且与其他组比较差异有统计学意义( P＜0.05)各组 TWIST1、ZEB1 mRNA水平比较差异无统计学意义( P＞0.05)。联合组可以显著抑制 Bcl?2、Caspase?3及 Survivin表达,上调,Bax表达,降低 Bcl?2、Bax比值,其效果明显高于吉西他滨组和大黄素组( P＜0.05)。联合组 E?cadhcrin蛋白表达显著高于其他组, P?gp、TWIST1、ZEB1蛋白表达显著低于其他组( P＜0.05)。结论大黄素能够下调 MDR?1降低胰腺癌细胞对吉西他滨耐药性,并能通过提高 miRNA?1271抑制胰腺癌细胞 EMT转化。     

青蒿琥酯增强吉西他滨的抗胰腺癌活性

目的 探讨青蒿琥酯对吉西他滨化疗胰腺癌的辅助治疗作用并研究其机制。方法 MTT法检测胰腺癌细胞系Capan-2在青蒿琥酯和不同浓度吉西他滨处理下的细胞活力。Western blot检测吉西他滨和青蒿琥酯对Capan-2细胞FOXO1、Noxa、Bim表达水平,细胞色素C、Smac/DIABLO从线粒体中的释放量及caspase-9、caspase-3活化水平的影响。流式细胞术检测Capan-2细胞的线粒体膜电位和凋亡率。结果 青蒿琥酯辅助治疗明显提高吉西他滨对Capan-2细胞的杀伤活性。青蒿琥酯处理能显著促进Capan-2细胞FOXO1的表达,转染FOXO1小干扰RNA （FOXO1 siRNA）后,青蒿琥酯对吉西他滨的辅助治疗效果受到明显抑制。青蒿琥酯联合吉西他滨能显著诱导Capan-2细胞中Noxa和Bim的过表达,线粒体膜电位的降低,细胞色素C、Smac/DIABLO的释放,caspase-9、caspase-3的活化及凋亡的发生。转染FOXO1siRNA后,青蒿琥酯联合吉西他滨对Capan-2细胞的凋亡诱导途径受到显著抑制。结论 青蒿琥酯可上调FOXO1的表达,增强吉西他滨对胰腺癌细胞的凋亡诱导活性。     

吉西他滨介导胰腺癌细胞三磷酸腺苷结合转运体G2质膜移位诱发耐药的研究

目的探讨三磷酸腺苷结合转运体G2(ABCG2)与胰腺癌采用吉西他滨化疗抵抗的相互关系。方法观察胰腺癌手术患者的癌和癌旁组织免疫组化及胰腺癌细胞系中ABCG2的表达,流式细胞术测定胰腺癌细胞系在吉西他滨刺激0、3、12、24、48、72 h后的ABCG2膜表达结果,选择高表达水平的PANC-1细胞株进行相关机制试验。观察吉西他滨作用0、15、30、45、120、180 min后P-AKT表达水平,及用PI3K/AKT抑制剂LY294002或AKT si RNA阻断AKT表达后ABCG2表达水平和生存率变化。免疫荧光共聚焦技术观察在对照、吉西他滨、LY294002和吉西他滨+LY294002 4种实验条件下细胞膜膜表面的ABCG2蛋白表达变化。结果 ABCG2蛋白表达量在胰腺癌组织中较癌旁组织明显增高,ABCG2+细胞在不同病理分化程度的胰腺癌细胞中均有表达。胰腺癌细胞系在吉西他滨刺激后总ABCG2水平无明显变化,而ABCG2+细胞数有明显增加。PANC-1在吉西他滨刺激后细胞总ABCG2蛋白表达无明显变化;p-AKT蛋白表达水平呈现增加趋势;ABCG2+细胞数明显增加,并呈时间相关性。LY294002或LY294002+吉西他滨处理细胞后ABCG2总蛋白表达无明显变化,si RNA-AKT2基因敲除或LY294002处理后,肿瘤细胞对吉西他滨的敏感性明显增加。免疫荧光共聚焦表明吉西他滨调节胰腺癌PI3K/AKT信号分子,介导ABCG2质膜移位。结论吉西他滨活化PI3K/AKT信号分子,促进ABCG2质膜移位,抑制胰腺癌化疗敏感性。     

Smac/DIABLO对胰腺癌SW1990细胞凋亡及化疗敏感性的影响

目的研究Smac/DIABLO与胰腺癌对TRAIL和吉西他滨化疗敏感性的关系。方法构建Flagpc3.1-Smac外源表达质粒,在SW1990细胞中过表达SMAC,采用实时定量荧光PCR方法检测Smac mRNA水平;应用Western免疫印迹法检测caspase-3、caspase-9和Bcl-2蛋白水平;利用MTT法检测转染Flag-pc3.1-Smac和TRAIL、吉西他滨处理后SW1990细胞增殖能力的改变;利用流式细胞术检测转染Flag-pc3.1-Smac后SW1990细胞的凋亡情况。结果转染Flag-pc3.1-Smac后,SW1990细胞中Smac mRNA和蛋白质表达水平显著高于对照组;MTT试验显示,过表达Flag-pc3.1-Smac的SW1990细胞在1~5 d的培养期中490 nm处吸光值均低于对照组。结论 Smac/DIABLO促进胰腺癌SW1990细胞凋亡,并增加对TRAIL和吉西他滨的化疗敏感性。     

Role of nuclear factor-kappaB on emodin-induced sensitization of pancreatic cancer to gemcitabine

In view of gemcitabine resistance has limited clinical activity of gemcitabine as a cellulotoxic drug in pancreatic cancer patients, this study is designed to investigate the effect of emodin on the sensitivity of pancreatic cancer to gemcitabine as well as its mechanism. After gemcitabine-resistant pancreatic cancer cell line (SW1990/GZ) was established by escalating doses of gemcitabine serially in pancreatic cancer cell line (SW1990). The cellular proliferation was detected by cell counting kit-8 (CCK-8) assay. Flow cytometry (FCM) was used to determine apoptosis of pancreatic cancer cells. The activity of NF-kappaB in pancreatic cancer cells was measured by electrophoretic mobility shift assay (EMSA). Western blotting was used to detect the protein expression of Bcl-2 and Survivin in SW1990/GZ cells. Metastatic model simulating human pancreatic cancer was established by orthotopic implantation of histologically intact human tumor tissue into pancreatic wall of nude mice. Also, immunohistochemistry was used to detect the positive expression of Ki-67, NF-kappaB, Bcl-2 and Survivin in the tumors. The results show that pretreatment of cells with emodin followed by gemcitabine induced a higher percentage of growth inhibition and apoptosis of pancreatic cancer cells than that of gemcitabine alone. In addition to in vitro results, emodin in combination with gemcitabine is much more effective as an antitumor agent compared to either agent alone in the orthotopic tumor model. Further study showed that the emodin with or without gemcitabine significantly down-regulates NF-kappaB and its regulated molecules such as Bcl-2 and Survivin proteins both in vitro and in vivo. It is concluded that inactivation of NF-kappaB signaling pathway by emodin resulting in the chemosensitization of pancreatic cancer to gemcitabine, which is likely to be an important and novel strategy for the treatment of pancreatic cancer.     

Gemcitabine combined with gum mastic causes potent growth inhibition and apoptosis of pancreatic cancer cells

Aim:

To investigate the antiproliferative and apoptotic effects of gemcitabine combined with gum mastic and the underlying mechanisms in human pancreatic cancer cell lines.

Methods:

Cell proliferation and apoptosis were examined using the methyl thiazolyl tetrazolium (MTT) assay and propidium iodine staining, respectively. The expression of Bcl-2, Bax, NF-κB p65 subunit, and IκBα protein was measured using Western blotting.

Results:

Gemcitabine 0.01−100 μg/mL inhibited cell proliferation and induced apoptosis in both pancreatic cancer BxPC-3 and COLO 357 cells. Gum mastic 40 μg/mL significantly potentiated the antiproliferative and apoptotic effects of gemcitabine 10 μg/mL after 72-h treatment. When cells were treated with gemcitabine in combination with gum mastic, the IκBα level was increased, whereas NF-κB activation was blocked; the expression of Bax protein was substantially increased, but Bcl-2 protein was down-regulated.

Conclusion:

Gemcitabine combined with gum mastic causes potent apoptosis in pancreatic cancer cells. The combination may be an effective therapeutic strategy for pancreatic cancer.     

UBE2M-mediated p27(Kip1) degradation in gemcitabine cytotoxicity

Gemcitabine (2'-deoxy-2', 2'-difluorocytidine; Gem) is a nucleoside anti-metabolite and is commonly used for treating various human cancers including human bladder carcinoma. Gemcitabine not only functions as a suicide nucleoside analog but also inhibits DNA polymerase activity and results in the termination of chain elongation. Using 2-dimensional gel electrophoresis analysis, a Gem-induced protein was identified as UBE2M (a.k.a. UBC12), a NEDD8 conjugation E2 enzyme which contributes to protein degradation. Gem induced UBE2M expression at both RNA and protein levels in several human cancer cell lines. The induction of UBE2M by Gem was accompanied by a reduction in p27(Kip1) protein levels, which could be restored by silencing UBE2M expression with siRNA or by treating cells with the proteasome inhibitor MG132, indicating that UBE2M mediates Gem-induced p27(Kip1) protein degradation. The induction of UBE2M and reduction of p27(Kip1) by Gem were prevented by the PI3K inhibitor LY294002. These results indicate that PI3K activity is necessary for Gem-induced UBE2M expression and that UBE2M facilitates degradation of p27(Kip1). Notably, silencing of UBE2M expression reduced Gem sensitivity in NTUB1 cells, suggesting that UBE2M mediates in part cell sensitivity to Gem, possibly by degradation of p27(Kip1). Analysis of Gem-resistant sub lines also showed that loss of UBE2M and increased p27(Kip1) expression were associated with the acquisition of drug resistance. In conclusion, our results demonstrate a role for UBE2M in mediating cytotoxicity of gemcitabine in human urothelial carcinoma cells while also suggesting a potential function of p27(Kip1) in drug resistance.     

UBE2M-mediated p27 degradation in gemcitabine cytotoxicity

Gemcitabine (2′-deoxy-2′, 2′-difluorocytidine; Gem) is a nucleoside anti-metabolite and is commonly used for treating various human cancers including human bladder carcinoma. Gemcitabine not only functions as a suicide nucleoside analog but also inhibits DNA polymerase activity and results in the termination of chain elongation. Using 2-dimensional gel electrophoresis analysis, a Gem-induced protein was identified as UBE2M (a.k.a. UBC12), a NEDD8 conjugation E2 enzyme which contributes to protein degradation. Gem induced UBE2M expression at both RNA and protein levels in several human cancer cell lines. The induction of UBE2M by Gem was accompanied by a reduction in p27^Kip1 protein levels, which could be restored by silencing UBE2M expression with siRNA or by treating cells with the proteasome inhibitor MG132, indicating that UBE2M mediates Gem-induced p27^Kip1 protein degradation. The induction of UBE2M and reduction of p27^Kip1 by Gem were prevented by the PI3K inhibitor LY294002. These results indicate that PI3K activity is necessary for Gem-induced UBE2M expression and that UBE2M facilitates degradation of p27^Kip1. Notably, silencing of UBE2M expression reduced Gem sensitivity in NTUB1 cells, suggesting that UBE2M mediates in part cell sensitivity to Gem, possibly by degradation of p27^Kip1. Analysis of Gem-resistant sub lines also showed that loss of UBE2M and increased p27^Kip1 expression were associated with the acquisition of drug resistance. In conclusion, our results demonstrate a role for UBE2M in mediating cytotoxicity of gemcitabine in human urothelial carcinoma cells while also suggesting a potential function of p27^Kip1 in drug resistance.     

Andrographolide causes apoptosis via inactivation of STAT3 and Akt and potentiates antitumor activity of gemcitabine in pancreatic cancer

Gemcitabine is a first-line drug utilised in the chemotherapy of pancreatic cancer; however, this drug induces chemo-resistance and toxicity to normal tissue during treatment. Here, we firstly report that andrographolide (ANDRO) alone not only has anti-pancreatic cancer activity, but it also potentiates the anti-tumour activity of gemcitabine. Treatment with ANDRO alone inhibits proliferation of the pancreatic cancer cell lines in a dose- and time-dependent manner in vitro. Interestingly, ANDRO induces cell cycle arrest and apoptosis of pancreatic cancer cells by inhibiting STAT3 and Akt activation, upregulating the expression of p21^WAF1 and Bax, and downregulating the expression of cyclinD1, cyclinE, survivin, X-IAP and Bcl-2. Additionally, ANDRO combined with gemcitabine significantly induce stronger cell cycle arrest and more obvious apoptosis than each single treatment. The mechanistic study demonstrates that this synergistic effect is also dependent on the inhibition of STAT3 and Akt activations which subsequently regulates the pathways involved in the apoptosis and cell cycle arrest. Furthermore, both ANDRO alone and the combination treatments exhibit efficacious anti-tumour activity in vivo. Overall, our results provide solid evidence supporting that ANDRO alone or its combination with gemcitabine is a potential chemotherapeutic approach for treating human pancreatic cancer in clinical practice.     

Allitridi induces apoptosis by affecting Bcl-2 expression and caspase-3 activity in human gastric cancer cells

INTRODUCTION Gastric cancer is the second leading cause of can-cer mortality in the world and is the leading cause ofcancer mortality in China[1]. Numerous epidemiologicalinvestigations have demonstrated that an inverse corre-lation between gastric cancer incidence and garlic con-sumption[2-8]. For example, Linqu County has gastriccancer rate that was 15 times higher than that of Cang-shan County in Shangdong Province[10], and the deathrate of gastric cancer in Linqu County was 7…     

6，8- 二- 三氟甲基-5- 羟基-7- 乙酰氧基白杨素的抗胃癌作用

目的研究6,8-二-三氟甲基-5-羟基-7-乙酰氧基白杨素（dFMAChR）的体外抗胃癌作用。方法体外培养人胃癌SGC-7901细胞。MTT比色法测定细胞活性。PI染色流式细胞术（FCM）分析细胞凋亡率。WesternBlot检测细胞NF-κB、Bcl-2、Bax蛋白表达。结果 dFMAChR显著抑制体外培养人胃腺癌SGC-7901细胞活性,呈剂量依赖性;dFMAChR有效诱导人胃腺癌SGC-7901细胞凋亡;dFMAChR下调NF-κB（p65）、Bcl-2蛋白表达,同时,上调Bax蛋白表达,呈浓度依赖性。结论 6,8-二-三氟甲基-5-羟基-7-乙酰氧基白杨素具有抗胃癌作用,其机制可能与其抑制NF-κB活化、下调Bcl-2蛋白表达和上调Bax蛋白表达相关。     

维生素C体外抑制宫颈癌H eLa细胞株生长及其机制的研究

目的探讨维生素C对人宫颈癌HeLa细胞生长和凋亡的影响,并初步探讨其机制。方法用MTT比色法检测细胞生长抑制作用,流式细胞术测定各组细胞周期分布和凋亡率,以间接免疫荧光法流式细胞术检测相关蛋白的表达,以TRAP-PCR-ELISA法测定端粒酶活性。结果维生素C能以浓度依赖和时间依赖方式阻滞HeLa细胞于G0/G1期,抑制其生长并诱导凋亡。同时HPV18 E6、Bc l-2蛋白表达下调,wtp53、Bax蛋白表达上调,端粒酶活性降低。结论维生素C可抑制HeLa细胞生长,使其细胞周期阻滞于G0/G1期,并诱导其凋亡;其原因在于病毒癌基因E6表达的降低,以及由此而引起的端粒酶活性的降低和细胞内蛋白表达水平的改变,包括HPV18 E6、Bc l-2蛋白表达的下调,wtp53、Bax蛋白表达的上调。