质子泵抑制剂提高人胃腺癌细胞化学治疗敏感性研究  

Pretreatment with proton pump inhibitor increases sensitivity of human gastric adenocarcinoma cell line to antitumor drugs

作　　者：陈敏[1] 邹晓平[1] 曹俊[1] 张斌[1] 刘文佳[1] 吴育美[1] 林海[1] 

机构地区：[1]南京大学医学院附属鼓楼医院消化科,210008

出　　处：《中华消化杂志》2009年第7期463-467,共5页Chinese Journal of Digestion

摘　　要：目的探讨质子泵抑制剂（PPI）泮托拉唑（PPZ）是否通过抑制空泡型质子泵来逆转细胞内外pH梯度，从而增加肿瘤细胞化学治疗敏感性，并探讨PPZ最佳预处理时间、剂量及机制。方法免疫印迹及免疫荧光法比较PPZ处理前、后人胃低分化腺癌细胞株SGC7901空泡型质子泵的表达及胞内分布变化。用BCECF-AM荧光探针检测不同浓度PPZ作用不同时间对细胞内pH值的影响。用四甲基偶氮唑盐比色法和膜联蛋白V-异硫氰酸荧光素-碘化丙啶试剂盒检测化学治疗药物联合PPZ的细胞毒性和细胞凋亡变化。用阿霉素检测PPZ对细胞内药物蓄积和潴留量的影响。结果10和100μg／m[PPZ作用24h后，空泡型质子泵表达的相对吸光度值（1．19±0．03和0．70±0．03）明显低于空白对照组（1．53±0．05），而1μg／ml PPZ可促进其表达（2．29±0．06）。在10μg／mlPPZ作用第6和12小时，可见其对空泡型质子泵表达的抑制作用（相对吸光度值分别为0．32±0．02和0．13±0．02），在作用24h后可改变空泡型质子泵的细胞内分布。10和100μg／ml PPZ作用24h可使细胞内pH值（7．44±0．09和7．31±0．06）明显低于空白对照组（7．51±0．05），10μg／ml及以上浓度的PPZ可逆转细胞内外pH梯度。PPZ预处理24h后予化学药物治疗的细胞存活率（58．71％±1．18％）低于单用化学治疗药物（74．33％±1．77％，P〈0．05），其总凋亡率（80．81％±1．16％）和早期凋亡率（77．52％±1．13％）显著高于单用化学治疗药物（26．42％±1．19％和23．18％±0．92％，P值均〈0．01）。20、50和100μg／ml PPZ作用24h后的阿霉素释放指数（0．164±0．013、0．162±0．015、0．152±0．012）低于空白对照组（0．277±0．011，P值均〈0．01）。结论PPZ预处理可提高人胃腺癌细胞的化学治疗敏感性。Objective To investigate whether pantoprazole （PPZ）, a proton pump inhibitor, could reverse the transmember pH gradient by inhibiting vacuolar H^＋-ATPase so as to increase the sensitivity of human gastric adenocarcinoma cell line SGC7901 to antitumor drugs and to evaluate the optimal time of drug administration, dosage of PPZ and the possible mechanism. Methods Western blotting and immunofluorescent staining were used to determine the expression and intracellular distribution of vacuolar H＋-ATPase in human gastric adenocarcinoma cell line SGC7901 with or without PPZ pretreatment. 2＇, 7＇-bis-（2-carboxyethyl）-5-（and-6 ）-carboxyfluorescein, acetoxymethyl ester （BCECF AM） fluorescent probe was used to measure the intracellular pH value of human gastric adenocarcinoma cell line SGC7901 which pretreated with different dose and time of PPL Methyl thiazolyl tetrazolium （MTT） assay and annexin V fluorescent isothiocyanate-propidium iodide double staining were performed to evaluate the cytotoxic effects and apoptosis of cells treated with ant＋tumor drugs combined with PPZ. Adriamycin （ADR） was used as probe to estimate drug accumulation and retention with PPZ pretreatment. Results After 24 hours, the expression of vacuolar H＋-ATPase in cells pretreated with PPZ of 10μg/ml （1.19±0.03） or 100 μg/ml （0. 70±0. 03） was significantly lower than that in blank control （1.53±0.05）, but this expression was increased by pretreatment with PPZ of 1 μg/ml （2.29±0.06,P〈0.05）. The inhibitory effects of PPZ （10 μg/ml） on vacuolar H＋-ATPase was observed at 6 hours （0.32±0.02）and 12 hours （0.13±0.02）. And it could alter the intracellular distribution of vacuolar H^＋-ATPase at 24-hours. The intracellular pH value in cells pretreated with PPZ of 10 μg/ml （7. 44±0. 09 ） or 100 μg/ml （7. 31±0. 06） was significantly decreased in comparison with untreated cells （7.51±0.05, P 〈 0.01 ）. After administration of antitumor drugs, the viability in SGC7901

关 键 词：空泡型质子泵 泮托拉唑 抗肿瘤剂 

分 类 号：R686[医药卫生—骨科学]