Reversal effects of pantoprazole on multidrug resistance in human gastric adenocarcinoma cells by down-regulating the V-ATPases/mTOR/HIF-1α/P-gp and MRP1 signaling pathway in vitro and in vivo.
Reversal effects of pantoprazole on multidrug resistance in human gastric adenocarcinoma cells by down-regulating the V-ATPases/mTOR/HIF-1α/P-gp and MRP1 signaling pathway in vitro and in vivo.
复制标题
DOI：
10.1002/jcb.24122
复制
发表时间：
2012-07
影响因子：
4
通讯作者：
Zou, Xiao-Ping
中科院分区：
文献类型：
--
作者：
Chen, Min;Huang, Shu-Ling;Zhang, Xiao-Qi;Zhang, Bin;Zhu, Hao;Yang, Vincent W.;Zou, Xiao-Ping
关键词：
探讨泮托拉唑（PPZ）对人胃腺癌细胞多药耐药（MDR）的逆转作用。将人胃腺癌细胞SGC 7901在37°C、5%CO2的潮湿气氛中在补充有10%胎牛血清和抗生素的RPMI-1640培养基中培养。阿霉素（ADR）耐药细胞在添加0.8 μg/ml ADR的情况下培养，维持MDR表型。方法：采用MTT法测定细胞内pH值，以ADR释放指数计算ADR释放指数，CCK-8法测定抗肿瘤药物的细胞毒性，BCECF-AM pH敏感荧光探针测定细胞内pH值，以培养基pH值作为细胞外pH值，MTT法测定细胞内pH值，MTT法测定细胞外pH值。采用Western blotting和免疫荧光染色分析方法检测液泡膜H+-ATPase的蛋白表达和胞内分布。ATPases（V-ATPases）、mTOR、HIF-1α、P-糖蛋白（P-gp）和多药耐药蛋白1（MRP 1）;将SGC 7901和SGC 7901/ADR细胞接种于无胸腺裸鼠。观察阿霉素对荷瘤小鼠肿瘤大小、重量的影响，并采用TUNEL法检测肿瘤组织中凋亡细胞的数量。当PPZ浓度大于20 μg/ml时，阿霉素释放指数降低，细胞内阿霉素浓度显著增加（P <0.01）。PPZ预处理可显著降低SGC 7901/ADR的ADR释放指数（P <0.01），并呈剂量依赖性。PPZ预处理也呈剂量依赖性地降低SGG 7901和SGC 7901/ADR的细胞活力。PPZ预处理24 h后，化疗药物对SGC 7901和SGC 7901/ADR细胞的杀伤作用最强（P < 0.05）。PPZ预处理组的阻力指数显著低于非PPZ预处理组（3.71 vs. 14.80）。PPZ浓度>10 μg/ml时，可显著降低SGC 7901和SGC 7901/ADR细胞的pHi，减小或逆转跨膜pH梯度（P < 0.05）。PPZ预处理可显著抑制亲本细胞和ADR耐药细胞V-ATP酶、mTOR、HIF-1α、P-gp和MRP 1的蛋白表达（P < 0.05）。体内实验进一步证实PPZ预处理能增强ADR对裸鼠移植瘤的抗肿瘤作用，并能提高移植瘤组织的凋亡指数。PPZ预处理可通过下调V-ATPases/mTOR/HIF-1α/P-gp和MRP 1信号通路增强抗肿瘤药物对SGC 7901的细胞毒作用，逆转SGC 7901/ADR的MDR。
To investigate reversal effects of pantoprazole (PPZ) on multidrug resistance (MDR) in human gastric adenocarcinoma cells in vivo and in vitro. Human gastric adenocarcinoma cell SGC7901 was cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and antibiotics in a humidified 5% CO2 atmosphere at 37°C. Adriamycin (ADR)-resistant cells were cultured with addition of 0.8 μg/ml of ADR maintaining MDR phenotype. ADR was used to calculate ADR releasing index; CCK-8 Assay was performed to evaluate the cytotoxicity of anti-tumor drugs; BCECF-AM pH-sensitive fluorescent probe was used to measure intracellular pH (pHi) value of cells, whereas pH value of medium was considered as extracellular pH (pHe) value; Western blotting and immunofluorescent staining analyses were employed to determine protein expressions and intracellular distributions of vacuolar H+-ATPases (V-ATPases), mTOR, HIF-1α, P-glycoprotein (P-gp), and multidrug resistant protein 1 (MRP1); SGC7901 and SGC7901/ADR cells were inoculated in athymic nude mice. Thereafter, effects of ADR with or without PPZ pretreatment were compared by determining the tumor size and weight, apoptotic cells in tumor tissues were detected by TUNEL assay. At concentrations greater than 20 μg/ml, PPZ pretreatment reduced ADR releasing index and significantly enhanced intracellular ADR concentration of SGC7901 (P <0.01). Similarly, PPZ pretreatment significantly decreased ADR releasing index of SGC7901/ADR dose-dependently (P <0.01). PPZ pretreatment also decreased cell viabilities of SGG7901 and SGC7901/ADR dose-dependently. After 24-h PPZ pretreatment, administration of chemotherapeutic agents demonstrated maximal cytotoxic effects on SGC7901 and SGC7901/ADR cells (P < 0.05). The resistance index in PPZ pretreatment group was significantly lower than that in non-PPZ pretreatment group (3.71 vs. 14.80). PPZ at concentration >10 μg/ml significantly decreased pHi in SGC7901 and SGC7901/ADR cells and diminished or reversed transmembrane pH gradient (P < 0.05). PPZ pretreatment also significantly inhibited protein expressions of V-ATPases, mTOR, HIF-1α, P-gp, and MRP1, and alter intracellular expressions in parent and ADR-resistant cells (P < 0.05). In vivo experiments further confirmed that PPZ pretreatment could enhance anti-tumor effects of ADR on xenografted tumor of nude mice and also improve the apoptotic index in xenografted tumor tissues. PPZ pretreatment enhances the cytotoxic effects of anti-tumor drugs on SGC7901 and reverse MDR of SGC7901/ADR by downregulating the V-ATPases/mTOR/HIF-1α/P-gp and MRP1 signaling pathway.
登录
查看更多内容
影响因子：
10.3
作者：
Luciani, F;Spada, M;Fais, S
通讯作者：
Fais, S
影响因子：
5.8
作者：
Anuchapreeda, S;Leechanachai, P;Limtrakul, P
通讯作者：
Limtrakul, P
影响因子：
4.8
作者：
LI, JF;EASTMAN, A
通讯作者：
EASTMAN, A
影响因子：
10.3
作者：
MARQUARDT, D;CENTER, MS
通讯作者：
CENTER, MS
大环双苄基衍生物逆转阿霉素耐药人骨髓性白血病（K562/A02）细胞中p-糖蛋白介导的多药耐药性
影响因子：
3.2
作者：
Li, Xia;Sun, Bin;Lou, Hong-Xiang
通讯作者：
Lou, Hong-Xiang