S100A9参与介导具核梭杆菌促结肠癌细胞的增殖与迁移的作用 *

目的:探讨S100A9在具核梭杆菌(Fusobacterium nucleatum,Fn)促结肠癌HCT116和SW480细胞增殖与迁移中的作用。方法:Fn感染HCT116和SW480细胞后,分别采用CCK8实验及Transwell实验检测结肠癌细胞的增殖及迁移能力的变化,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化;用S100A9 siRNA转染HCT116和SW480细胞后感染Fn,采用CCK8实验及Transwell实验分别检测两株细胞增殖及迁移能力的变化;利用NF-κB抑制剂BAY 11-7082预处理HCT116和SW480细胞后再感染Fn,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化。结果:(1)Fn可促进HCT116和SW480细胞增殖及迁移(P<0.001); (2)Fn感染的HCT116和SW480细胞中S100A9基因及蛋白质水平均较相应对照组明显升高,差异均具有高度显著性(P<0.01或P<0.001),即Fn上调S100A9的表达;(3)用S100A9 siRNA下调HCT116和SW480细胞中S100A9表达后,Fn促进这两株细胞增殖及迁移的作用则明显减弱,差异均具有高度显著性(P<0.001),表明S100A9参与介导Fn的促HCT116和SW480细胞增殖及迁移的作用; (4)在NF-κB通路抑制剂BAY 11-7082预处理的HCT116和SW480细胞组,Fn上调S100A9基因及蛋白质水平的作用明显被逆转,差异均具有高度显著性(P<0.01或P<0.001),表明激活NF-κB转录活性是Fn上调S100A9的机制之一。结论:Fn可上调S100A9表达且与NF-κB活化有关,上调S100A9是Fn促结肠癌细胞增殖及迁移的机制之一。     

S100A9参与介导具核梭杆菌促结肠癌细胞的增殖与迁移的作用

目的:探讨S100A9在具核梭杆菌(Fusobacterium nucleatum,Fn)促结肠癌HCT116和SW480细胞增殖与迁移中的作用。方法:Fn感染HCT116和SW480细胞后,分别采用CCK8实验及Transwell实验检测结肠癌细胞的增殖及迁移能力的变化,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化;用S100A9 siRNA转染HCT116和SW480细胞后感染Fn,采用CCK8实验及Transwell实验分别检测两株细胞增殖及迁移能力的变化;利用NF-κB抑制剂BAY 11-7082预处理HCT116和SW480细胞后再感染Fn,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化。结果:(1) Fn可促进HCT116和SW480细胞增殖及迁移(P 0.001);(2) Fn感染的HCT116和SW480细胞中S100A9基因及蛋白质水平均较相应对照组明显升高,差异均具有高度显著性(P 0. 01或P 0. 001),即Fn上调S100A9的表达;(3)用S100A9 siRNA下调HCT116和SW480细胞中S100A9表达后,Fn促进这两株细胞增殖及迁移的作用则明显减弱,差异均具有高度显著性(P 0. 001),表明S100A9参与介导Fn的促HCT116和SW480细胞增殖及迁移的作用;(4)在NF-κB通路抑制剂BAY 11-7082预处理的HCT116和SW480细胞组,Fn上调S100A9基因及蛋白质水平的作用明显被逆转,差异均具有高度显著性(P 0. 01或P 0. 001),表明激活NF-κB转录活性是Fn上调S100A9的机制之一。结论:Fn可上调S100A9表达且与NF-κB活化有关,上调S100A9是Fn促结肠癌细胞增殖及迁移的机制之一。     

乙肝病毒X蛋白通过NF-κB通路调节肝癌细胞迁移、侵袭的研究

慢性乙肝病毒感染是亚洲国家肝癌的常见病因,乙肝病毒所表达的乙肝病毒X蛋白(Hepatitis B virus X protein,HBx)是肝癌发生发展的重要推动因子。核因子-κB(Nuclear factorκB,NF-κB)是模式识别受体下游重要的转录因子,肝细胞生长因子(Hepatocyte growth factor,HGF)/c-Met途径能够促进NF-κB活化,并通过活化的NF-κB来调节肝癌细胞的迁移、侵袭等生物学环节。但HBx是否直接通过NF-κB通路来调节肝癌细胞的迁移、侵袭尚未明确。为探究HBx通过NF-κB通路调节肝癌细胞迁移、侵袭的作用,本研究采用培养肝癌HepG2细胞并分组,空白对照组用不含药物及质粒的DMEM处理,空白质粒组转染1.2μg的pcDNA3.1空白质粒,HBx质粒组转染不同浓度的HBx表达质粒pcDNA3.1-HBx,HBx+PDTC组转染1.2μg的HBx表达质粒pcDNA3.1-HBx并用含有50μmol/L PDTC的DMEM进行处理;皮下注射HepG2细胞建立移植瘤小鼠模型,称量移植瘤的质量。检测细胞迁移及侵袭活力、细胞及移植瘤中NF-κB通路分子、迁移基因、侵袭基因的表达。结果显示,与空白对照组、空白质粒组比较,HBx质粒组细胞中NF-κB、HGF、c-Met、N-cadherin、Vimentin、MMP2、MMP9的蛋白表达水平及相对愈合面积、侵袭数目均明显增多(P0.05);与HBx质粒组比较,HBx+PDTC组细胞中NF-κB、HGF、c-Met、N-cadherin、Vimentin、MMP2、MMP9的蛋白表达水平及相对愈合面积、侵袭数目均明显减少(P0.05);与空白对照组、空白质粒组比较,HBx质粒组移植瘤的质量及移植瘤中NF-κB、HGF、c-Met的蛋白表达水平明显增加(P0.05)。本研究得出结论,HBx能够促进肝癌细胞的迁移、侵袭,且该作用与激活NF-κB通路有关。     

乙肝病毒X 蛋白通过激活NF-κB 信号通路上调ABCG2 的表达

目的:研究乙肝病毒X蛋白(HBx)通过核因子-κB(NF-κB)信号通路对半转运蛋白(ABCG2)的调节作用。方法:用特异性的NF-κB信号通路阻断剂PDTC阻断NF-κB信号通路,荧光双标激光扫描共聚焦显微镜观察L02细胞系转染HBx基因前后及PDTC加入前后NF-κB信号通路的激活、失活情况,同时用Real-time PCR和Western Blot技术检测转染前后及PDTC加入前后ABCG2在mRNA及蛋白水平的表达变化。结果:以L02细胞为参照,转染HBx基因后的L02-HBx细胞NF-κB信号通路被激活,ABCG2 mRNA和蛋白水平分别增加3.62±0.15和4.61±0.73倍,差异有统计学意义(P<0.05);PDTC作用24h后L02/HBx细胞NF-κB信号通路阻断,ABCG2 mRNA和蛋白表达分别为2.15±0.32倍和2.37±0.55倍,与未加入PDTC作用的L02-HBx细胞相比均有统计学意义(P<0.05)。结论:NF-κB信号通路是HBx上调ABCG2表达的途径之一。     

乙肝病毒X蛋白通过激活NF-κB信号通路上调ABCG2的表达

目的：研究乙肝病毒X蛋白（HBx）通过核因子-κB（NF-κB）信号通路对半转运蛋白（ABCG2）的调节作用。方法：用特异性的NF-κB信号通路阻断剂PDTC阻断NF-κB信号通路,荧光双标激光扫描共聚焦显微镜观察L02细胞系转染HBx基因前后及PDTC加入前后NF-κB信号通路的激活、失活情况,同时用Real-time PCR和Western Blot技术检测转染前后及PDTC加入前后ABCG2在mRNA及蛋白水平的表达变化。结果：以L02细胞为参照,转染HBx基因后的L02-HBx细胞NF-κB信号通路被激活,ABCG2 mRNA和蛋白水平分别增加3.62±0.15和4.61±0.73倍,差异有统计学意义（P〈0.05）;PDTC作用24h后L02/HBx细胞NF-κB信号通路阻断,ABCG2 mRNA和蛋白表达分别为2.15±0.32倍和2.37±0.55倍,与未加入PDTC作用的L02-HBx细胞相比均有统计学意义（P〈0.05）。结论：NF-κB信号通路是HBx上调ABCG2表达的途径之一。     

S100A9对宫颈癌Hela细胞的增殖和迁移的影响及其机制探讨

为初步探讨S100A9在宫颈癌中的生物学作用,利用携带有人S100A9基因的腺病毒(Adh S100A9)感染于低表达S100A9的宫颈癌Hela细胞,通过MTT法检测细胞增殖能力,划痕愈合试验和Transwell试验检测细胞迁移能力,通过倒置显微镜观察细胞形态变化,采用RT-PCR和蛋白质印迹法检测上皮细胞标志蛋白E-钙黏着蛋白(E-cadherin,E-cad)、间质细胞标志蛋白波形蛋白(vimentin,Vim)及Wnt/β-联蛋白(β-catenin,β-cat)信号通路相关分子的表达。结果显示,与对照组相比,过表达S100A9的Hela细胞增殖活性增强、迁移率增高,细胞形态由"铺路石"样向"梭形"转变,排列紊乱,伴随E-cad降低(P0.05)和Vim升高(P0.01);同时,过表达S100A9的Hela细胞中Wnt/β-cat信号通路的关键分子β-联蛋白水平增加(P0.01),并且入核增多(P0.01),该通路的下游靶基因c-myc、Snail及Twist表达也明显上调。该研究结果提示,S100A9促进宫颈癌Hela细胞的增殖、迁移和上皮–间质转化(epithelial-mesenchymal transition,EMT),激活Wnt/β-cat信号通路;S100A9促进增殖和迁移作用的机制可能与其促进EMT和激活Wnt/β-cat信号通路相关。     

RNA干扰CD151基因表达抑制肝癌细胞侵袭

目的研究RNA干扰(RNA interference RNAi)抑制CD151表达对人类肝癌细胞迁移侵袭的影响及分子机制。方法将CD151-siRNA在脂质体介导下瞬时转入人肝癌HepG2细胞,倒置荧光显微镜观察转染效率,用qPCR,western blot检测HepG2细胞CD151mRNA和蛋白表达,体外研究肿瘤细胞迁移和侵袭能力,并检测相关信号通路的变化。结果成功转染CD151-siRNA后,HepG2细胞CD151基因的表达与正常对照组和阴性对照组相比,mRNA和蛋白表达水平明显降低(P0.05),细胞迁移和侵袭能力明显下降(P0.05),同时,沉默CD151的表达,FAK,ERK的磷酸化受抑制。结论CD151-siRNA能有效抑制人肝癌细胞CD151基因mRNA和蛋白的表达,通过抑制FAK,ERK蛋白的磷酸化水平,降低细胞的迁移和侵袭力。     

新的蒽醌类衍生物1-硝基-2-酰基蒽醌-苯丙氨酸通过抑制NF-κB/p65信号通路降低乳腺癌MCF-7细胞的迁移能力北大核心CSCD

肿瘤细胞的侵袭和转移与大多数癌症患者的死亡率密切相关。了解肿瘤细胞的迁移机制可为阻断肿瘤细胞的转移提供一个关键的策略。蒽醌衍生物具有一定的抗肿瘤作用,我们结合抗肿瘤药物的作用机制以及蒽醌类衍生物的构效关系,设计合成了一类新的酰胺蒽醌衍生物1-硝基-2-酰基蒽醌-苯丙氨酸(简称C7),发现其具有很好的抗肿瘤活性。为了探究蒽醌类衍生物C7对人乳腺癌MCF-7细胞迁移的作用及其机制,本文首先采用MTT比色法检测蒽醌类衍生物C7对人乳腺癌细胞MCF-7生长活力的影响,结果证明较高浓度(60~100μg/m L)的蒽醌类衍生物C7对乳腺癌MCF-7细胞的增殖具有明显的抑制作用。其次,采用细胞划痕实验检测C7对MCF-7细胞迁移的影响,发现较低浓度(20~40μg/m L)的蒽醌类衍生物C7可以显著降低MCF-7细胞的迁移率。为进一步探究C7抑制MCF-7细胞迁移的分子机制,通过免疫荧光技术检测NF-κB/p65蛋白的核转位情况;同时利用qRT-PCR及Western印迹实验检测C7对MCF-7细胞中NF-κB/p65通路及迁移相关基因和蛋白质表达的影响。结果表明C7可以下调细胞质中IκBα的磷酸化,降低NF-κB/p65蛋白的核转位,减小MMP-2和MMP-9蛋白的表达。因此,C7可能是通过抑制NF-κB/p65信号通路的活化,抑制MMP-2和MMP-9蛋白的表达,进而抑制MCF-7细胞的迁移。     

NF-κB抑制剂通过逆转LPS诱导的BMPRⅡ下调改善肺血管重构

动脉性肺动脉高压(pulmonary arterial hypertension, PAH)的发生、发展与骨形态发生蛋白受体II型(bone morphogenetic protein receptor type II, BMPRII)编码基因的遗传突变和核因子κB (nuclear factorκB, NF-κB)通路介导的炎症反应密切相关。本文旨在研究NF-κB通路抑制剂对脂多糖(lipopolysaccharide, LPS)诱导的肺动脉内皮细胞损伤的作用。用1μg/mL的LPS处理人肺动脉内皮细胞,用免疫印迹和qPCR检测BMPRII和白介素8 (interleukin-8, IL-8)的表达水平。腹腔注射野百合碱(monocrotaline, MCT)建立大鼠PAH模型,用免疫荧光染色法检测肺动脉内皮细胞BMPRII和IL-8的表达情况,检测模型大鼠心脏血流动力学变化和肺血管重构情况。结果显示,LPS可引起人肺动脉内皮细胞BMPRII的表达下调和IL-8的表达上调,NF-κB抑制剂BAY11-7082 (10μmol/L)可逆转LPS的作用。在MCT-PAH大鼠模型中,肺动脉内皮细胞BMPRII表达下调,IL-8表达上调,右心室/(左心室+室间隔)重量比值[weight ratio of right ventricle to left ventricle plus septum, RV/(LV+S)]和右心室收缩压(right ventricular systolic pressure, RVSP)显著升高,心输出量(cardiac output, CO)和三尖瓣环收缩期位移(tricuspid annular plane systolic excursion, TAPSE)明显降低,肺血管壁明显增厚,连续21天腹腔注射BAY11-7082 (5 mg/kg)可逆转上述变化。以上结果提示,LPS通过NF-κB信号通路下调BMPRII的表达水平,促进PAH的发生、发展,因此NF-κB信号通路可作为PAH潜在治疗靶点。     

新的蒽醌类衍生物1-硝基-2-酰基蒽醌-苯丙氨酸通过抑制NF-κB/p65信号通路降低乳腺癌MCF-7细胞的迁移能力

肿瘤细胞的侵袭和转移与大多数癌症患者的死亡率密切相关。了解肿瘤细胞的迁移机制可为阻断肿瘤细胞的转移提供一个关键的策略。蒽醌衍生物具有一定的抗肿瘤作用,我们结合抗肿瘤药物的作用机制以及蒽醌类衍生物的构效关系,设计合成了一类新的酰胺蒽醌衍生物1-硝基-2-酰基蒽醌-苯丙氨酸(简称C7),发现其具有很好的抗肿瘤活性。为了探究蒽醌类衍生物C7对人乳腺癌MCF-7细胞迁移的作用及其机制,本文首先采用MTT比色法检测蒽醌类衍生物C7对人乳腺癌细胞MCF-7生长活力的影响,结果证明较高浓度(60~100μg/m L)的蒽醌类衍生物C7对乳腺癌MCF-7细胞的增殖具有明显的抑制作用。其次,采用细胞划痕实验检测C7对MCF-7细胞迁移的影响,发现较低浓度(20~40μg/m L)的蒽醌类衍生物C7可以显著降低MCF-7细胞的迁移率。为进一步探究C7抑制MCF-7细胞迁移的分子机制,通过免疫荧光技术检测NF-κB/p65蛋白的核转位情况;同时利用qRT-PCR及Western印迹实验检测C7对MCF-7细胞中NF-κB/p65通路及迁移相关基因和蛋白质表达的影响。结果表明C7可以下调细胞质中IκBα的磷酸化,降低NF-κB/p65蛋白的核转位,减小MMP-2和MMP-9蛋白的表达。因此,C7可能是通过抑制NF-κB/p65信号通路的活化,抑制MMP-2和MMP-9蛋白的表达,进而抑制MCF-7细胞的迁移。     

Hepatitis B virus X protein drives multiple cross-talk cascade loops involving NF-κB, 5-LOX, OPN and Capn4 to promote cell migration

Hepatitis B virus X protein (HBx) plays an important role in the development of hepatocellular carcinoma (HCC). However, the mechanism remains unclear. Recently, we have reported that HBx promotes hepatoma cell migration through the upregulation of calpain small subunit 1 (Capn4). In addition, several reports have revealed that osteopontin (OPN) plays important roles in tumor cell migration. In this study, we investigated the signaling pathways involving the promotion of cell migration mediated by HBx. We report that HBx stimulates several factors in a network manner to promote hepatoma cell migration. We showed that HBx was able to upregulate the expression of osteopontin (OPN) through 5-lipoxygenase (5-LOX) in HepG2-X/H7402-X (stable HBx-transfected cells) cells. Furthermore, we identified that HBx could increase the expression of 5-LOX through nuclear factor-κB (NF-κB). We also found that OPN could upregulate Capn4 through NF-κB. Interestingly, we showed that Capn4 was able to upregulate OPN through NF-κB in a positive feedback manner, suggesting that the OPN and Capn4 proteins involving cell migration affect each other in a network through NF-κB. Importantly, NF-κB plays a crucial role in the regulation of 5-LOX, OPN and Capn4. Thus, we conclude that HBx drives multiple cross-talk cascade loops involving NF-κB, 5-LOX, OPN and Capn4 to promote cell migration. This finding provides new insight into the mechanism involving the promotion of cell migration by HBx.     

RPS3a over-expressed in HBV-associated hepatocellular carcinoma enhances the HBx-induced NF-κB signaling via its novel chaperoning function

Hepatitis B virus (HBV) infection is one of the major causes of hepatocellular carcinoma (HCC) development. Hepatitis B virus X protein (HBx) is known to play a key role in the development of hepatocellular carcinoma (HCC). Several cellular proteins have been reported to be over-expressed in HBV-associated HCC tissues, but their role in the HBV-mediated oncogenesis remains largely unknown. Here, we explored the effect of the over-expressed cellular protein, a ribosomal protein S3a (RPS3a), on the HBx-induced NF-κB signaling as a critical step for HCC development. The enhancement of HBx-induced NF-κB signaling by RPS3a was investigated by its ability to translocate NF-κB (p65) into the nucleus and the knock-down analysis of RPS3a. Notably, further study revealed that the enhancement of NF-κB by RPS3a is mediated by its novel chaperoning activity toward physiological HBx. The over-expression of RPS3a significantly increased the solubility of highly aggregation-prone HBx. This chaperoning function of RPS3a for HBx is closely correlated with the enhanced NF-κB activity by RPS3a. In addition, the mutational study of RPS3a showed that its N-terminal domain (1-50 amino acids) is important for the chaperoning function and interaction with HBx. The results suggest that RPS3a, via extra-ribosomal chaperoning function for HBx, contributes to virally induced oncogenesis by enhancing HBx-induced NF-κB signaling pathway.     

S100A8 and S100A9 promotes invasion and migration through p38 mitogen-activated protein kinase-dependent NF-κB activation in gastric cancer cells

S100A8 and S100A9 (S100A8/A9) are low-molecular weight members of the S100 family of calcium-binding proteins. Recent studies have reported S100A8/A9 promote tumorigenesis. We have previously reported that S100A8/A9 is mostly expressed in stromal cells and inflammatory cells between gastric tumor cells. However, the role of environmental S100A8/A9 in gastric cancer has not been defined. We observed in the present study the effect of S100A8/A9 on migration and invasion of gastric cancer cells. S100A8/ A9 treatment increased migration and invasionat lower concentrations that did not affect cell proliferation and cell viability. S100A8/A9 caused activation of p38 mitogenactivated protein kinase (MAPK) and nuclear factor-κB (NF-κB). The phosphorylation of p38 MAPK was not affected by the NF-κB inhibitor Bay whereas activation of NF-κB was blocked by p38 MAPK inhibitor SB203580, indicating that S100A8/A9-induced NF-κB activation is mediated by phosphorylation of p38 MAPK. S100A8/A9-induced cell migration and invasion was inhibited by SB203580 and Bay, suggesting that activation of p38 MAPK and NF-κB is involved in the S100A8/A9 induced cell migration and invasion. S100A8/A9 caused an increase in matrix metalloproteinase 2 (MMP2) and MMP12 expression, which were inhibited by SB203580 and Bay. S100A8/A9-induced cell migration and invasion was inhibited by MMP2 siRNA and MMP12 siRNA, indicating that MMP2 and MMP12 is related to the S100A8/A9 induced cell migration and invasion. Taken together, these results suggest that S100A8/A9 promotes cell migration and invasion through p38 MAPKdependent NF-κB activation leading to an increase of MMP2 and MMP12 in gastric cancer.     

Protein profile in HBx transfected cells: A comparative iTRAQ-coupled 2D LC-MS/MS analysis

The x protein of HBV (HBx) has been involved in the development of hepatocellular carcinoma (HCC), with a possible link to individual genotypes. Nevertheless, the underlying mechanism remains obscure. In this study, we aim to identify the HBx-induced protein profile in HepG2 cells by LC-MS/MS proteomics analysis. Our results indicated that proteins were differentially expressed in HepG2 cells transfected by HBx of various genotypes. Proteins associated with cytoskeleton were found to be either up-regulated (MACF1, HMGB1, Annexin A2) or down-regulated (Lamin A/C). These may in turn result in the decrease of focal adhesion and increase of cell migration in response to HBx. Levels of other cellular proteins with reported impact on the function of extracellular matrix (ECM) proteins and cell migration, including Ca²⁺-binding proteins (S100A11, S100A6, and S100A4) and proteasome protein (PSMA3), were affected by HBx. The differential protein profile identified in this study was also supported by our functional assay which indicated that cell migration was enhanced by HBx. Our preliminary study provided a new platform to establish a comprehensive cellular protein profile by LC-MS/MS proteomics analysis. Further downstream functional assays, including our reported cell migration assay, should provide new insights in the association between HCC and HBx.     

The X protein of hepatitis B virus inhibits apoptosis in hepatoma cells through enhancing the methionine adenosyltransferase 2A gene expression and reducing S-adenosylmethionine production

The X protein (HBx) of hepatitis B virus (HBV) is involved in the development of hepatocellular carcinoma (HCC), and methionine adenosyltransferase 2A (MAT2A) promotes the growth of liver cancer cells through altering S-adenosylmethionine homeostasis. Thus, we speculated that a link between HBx and MAT2A may contribute to HCC development. In this study, the effects of HBx on MAT2A expression and cell apoptosis were investigated, and the molecular mechanism by which HBx and MAT2A regulate tumorigenesis was evaluated. Results from immunohistochemistry analyses of 37 pairs of HBV-associated liver cancer tissues/corresponding peritumor tissues showed that HBx and MAT2A are highly expressed in most liver tumor tissues. Our in vitro results revealed that HBx activates MAT2A expression in a dose-dependent manner in hepatoma cells, and such regulation requires the cis-regulatory elements NF-κB and CREB on the MAT2A gene promoter. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) further demonstrated that HBx facilitates the binding of NF-κB and CREB to MAT2A gene promoter. In addition, overexpression of HBx or MAT2A inhibits cell apoptosis, whereas knockdown of MAT2A expression stimulates apoptosis in hepatoma cells. Furthermore, we demonstrated that HBx reduces MAT1A expression and AdoMet production but enhances MAT2β expression. Thus, we proposed that HBx activates MAT2A expression through NF-κB and CREB signaling pathways to reduce AdoMet production, inhibit hepatoma cell apoptosis, and perhaps enhance HCC development. These findings should provide new insights into our understanding how the molecular mechanisms underline the effects of HBV infection on the production of MAT2A and the development of HCC.     

HBx induces HepG-2 cells autophagy through PI3K/Akt�CmTOR pathway

Chronic hepatitis B virus infection is the dominant global cause of hepatocellular carcinoma (HCC), especially hepatitis B virus-X (HBx) plays a major role in this process. HBx protein promotes cell cycle progression, inactivates negative growth regulators, and binds to and inhibits the expression of p53 tumor suppressor gene and other tumor suppressor genes and senescence-related factors. However, the relationship between HBx and autophagy during the HCC development is poorly known. Previous studies found that autophagy functions as a survival mechanism in liver cancer cells. We suggest that autophagy plays a possible role in the pathogenesis of HBx-induced HCC. The present study showed that HBx transfection brought about an increase in the formation of autophagosomes and autolysosomes. Microtubule-associated protein light chain 3, Beclin 1, and lysosome-associated membrane protein 2a were up-regulated after HBx transfection. HBx-induced increase in the autophagic level was increased by mTOR inhibitor rapamycin and was blocked by treatment with the PI3K?CAkt inhibitor LY294002. The same results can also be found in HepG2.2.15 cells. These results suggest that HBx activates the autophagic lysosome pathway in HepG-2 cells through the PI3K?CAkt?CmTOR pathway.