摘要
目的 探讨蛋白激酶C(proteinkinaseC ,PKC)的激活剂及抑制剂对胃癌耐药细胞系P 糖蛋白 (P glycoprotein ,Pgp)功能及表达的影响。 方法 用免疫细胞化学及流式细胞仪方法检测Pgp在胃癌耐药细胞系SGC790 1/VCR及其药敏细胞系SGC790 1的表达 ,并观察了PKC的激活剂佛波酯 (phor bol 12 myristate 13 acctate ,PMA)和抑制剂H 7作用不同时间Pgp表达的变化。用免疫荧光双标记及共聚焦显微镜技术观察PKC α及Pgp在胃癌耐药细胞系及其药敏细胞系的共同表达。用罗丹明 12 3(Rohdamin 12 3,R12 3)作为荧光探针检测PMA及H 7对细胞内药物浓度的影响。结果 Pgp在耐药细胞系SGC790 1/VCR表达呈阳性 ,在药敏细胞系SGC790 1表达呈弱阳性。PKC α与Pgp在SGC790 1/VCR及SGC790 1细胞系细胞膜均有共同表达 ,以SGC790 1/VCR细胞系共同表达明显。PMA处理细胞 ,细胞内R12 3的平均荧光强度明显减低 ,Pgp的功能增强而表达减少 ,且有时间依赖性 ;H 7处理细胞 ,细胞内R12 3的平均荧光强度明显增加 ,Pgp的功能减弱而表达增加 ,也有时间依赖性。 结论 PKC可能通过调节Pgp的功能与表达而参与胃癌细胞多药耐药的形成。
Objective To study the effects of activation and inhibition agents of protein kinase C on the function and expression of P glycoprotein in drug resistant gastric cancer cells. Methods Immunohistochemical staining and FACS were used to determine the expression of P glycoprotein on drug resistant gastric cancer cells SGC7901/VCR and its parent cells SGC7901, which were treated or not treated by activation or inhibition agents of protein kinase C(PMA or H 7). Double labeled immunoflurescent and laser scan confocal microscopy were used to detect the coexpression of protein kinase C α and P glycoprotein on drug resistant gastric cancer cells. Rohdamin 123 was used as fluorescent probe to observe the effects of PMA and H 7 on the function of P glycoprotein and drug accumulation in SGC7901/VCR. Results P glycoprotein had positive staining on SGC7901 cells and much stronger staining on SGC7901/VCR cells. Protein kinase C α and P glycoprotein exhibited coexpression on SGC7901 cells and much higher coexpression level on SGC7901/VCR cells. When treated by PMA, SGC7901/VCR cells showed time dependent decreasing of drug accumulation, enhancing function and decreasing expression of P glycoprotein. When treated by H 7, SGC7901/VCR cells showed time dependent increasing of drug accumulation, weakening function and increasing expression of P glycoprotein.Conclusion Protein kinase C may play a role in multidrug resistance of gastric cancer cells via modulating the expression and function of P glycoprotein.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2001年第6期349-352,共4页
Chinese Journal of Digestion
基金
国家自然科学基金重点项目! (30 0 30 14 0 )
国家自然科学基金资助项目! (30 0 0 0 0 6 6 )
关键词
蛋白激酶C
多药耐药
P-糖蛋白
胃癌
Protein kinase C
Multidrug resistance
P glycoprotein
Gastric cancer
