蛋白激酶C抑制剂对宫颈癌细胞化疗药物耐药的影响

摘要

目的：观察蛋白激酶C抑制剂星形孢菌素对经紫杉醇处理的宫颈癌细胞增殖、凋亡及蛋白激酶C、P-糖蛋白（ P-gp）表达的变化，探讨蛋白激酶C抑制剂对宫颈癌细胞紫杉醇耐药的影响。方法培养人宫颈癌Hela细胞，设立星形孢菌素组、紫杉醇组、联合组及对照组。星形孢菌素组加入星形孢菌素0．6μmol/L；紫杉醇组加入紫杉醇0．1μmol/L；联合组先加入星形孢菌素0．6μmol/L，随后加入紫杉醇0．1μmol/L；对照组不加入星形孢菌素及紫杉醇。四组细胞干预后继续培养48 h，采用MTT法检测细胞增殖能力，流式细胞术检测细胞凋亡率，Western blot法检测蛋白激酶C及P-gp蛋白表达。结果星形孢菌素组、紫杉醇组及联合组增殖抑制率分别为38．11％±0．70％、49．96％±1．60％及60．31％±1．80％，联合组增殖抑制率高于星形孢菌素组及紫杉醇组（P均＜0．05），星形孢菌素组增殖抑制率低于紫杉醇组（ P＜0．05）。对照组、星形孢菌素组、紫杉醇组及联合组细胞凋亡率分别为7．13％±0．56％、17．73％±0．59％、36．51％±0．34％、56．72％±0．68％，星形孢菌素组、紫杉醇组及联合组细胞凋亡率均高于对照组（P均＜0．05），联合组细胞凋亡率高于其他三组（P均＜0．05）。星形孢菌素组、联合组蛋白激酶C及P-gp蛋白表达均低于对照组（P均＜0．05），联合组蛋白激酶C及P-gp蛋白表达均低于星形孢菌素组、紫杉醇组（P均＜0．01）。结论星形孢菌素可抑制经紫杉醇处理后的宫颈癌细胞的增殖、促进其凋亡，改善宫颈癌细胞对紫杉醇的耐药性。星形孢菌素可能通过减少P-gp蛋白表达改善宫颈癌细胞对紫杉醇的耐药性。%Objective To study the effect of protein kinase C inhibitors combined with paclitaxel on proliferation, apopto-sis, kinase C and P-glycoprotein (P-gp) expression of cervical cancer cell, in order to explore the effect of protein kinase C in-hibitor on the drug resistance of cervical cancer.Methods Cervical cancer Hela cells were cultured and divided into 4 groups, which were staurosporin group, paclitaxel group, combination group and control group.Staurosporin group were cultured with 0.6μmol/L staurosporin.Paclitaxel group were cultured with 0.1 μmol/L paclitaxel.Combination group were cultured with 0.6μmol/L staurosporin and 0.1μmol/L paclitaxel.Control group did not culture with staurosporin and paclitaxel.At 48 hours lat-er, the cell proliferation of each group was performed by MTT method;the cell apoptosis were detected by flow cytometry(FCM) method;the expression of protein kinase C and P-gp were detected by western-blot method.Results The proliferation of stau-rosporin group, paclitaxel group, combination group was 38.11%±0.70%, 49.96%±1.60%and 60.31%±1.80%, respec-tively.The proliferation of combination group was higher than that of staurosporin group and paclitaxel group(P<0.05).The proliferation of staurosporin group was lower than that of paclitaxel group(P<0.05).The cell apoptosis of control group, stauros-porin group, paclitaxel group and combination group was 7.13%±0.56%, 17.73%±0.59%, 36.51%±0.34%and 56.72%± 0.68%, respectively.The cell apoptosis of staurosporin group, paclitaxel group and combination group was higher than that of control group(P<0.05).The expression of protein kinase C and P-gp in staurosporin group and combination group were lower than those of control group(P<0.05).The expression of protein kinase C and P-gp in combination group were lower than those of staurosporin group and paclitaxel group(P<0.05).Conclusions The protein kinase C inhibits staurosporin can inhibit the proliferation of cervical cancer cells as well as induce its apoptosis, which means that staurosporin can improve the drug resist-ance of cervical cancer cells to paclitaxel.Staurosporin may invert the drug resistance in cervical cancer by reducing P-gp protein expression.