摘要
目的探讨姜黄素逆转非小细胞肺癌(NSCLC)酪氨酸激酶抑制剂(TKI)靶向药物耐药的分子机制。方法选用非小细胞肺癌NCI-H1975细胞,分别采用相应药物进行处理。采用MTT法检测姜黄素对NCI-H1975细胞增殖的抑制作用,同时检测姜黄素对NCI-H1975细胞耐药性的逆转作用,采用流式细胞术检测姜黄素对NCIH1975细胞凋亡率的影响,采用Western Blot法检测各组NCI-H1975细胞p-PI3K、p-Akt、p-Ras、p-ERK蛋白表达水平。结果与对照组比较,姜黄素5μmol/L组、10μmol/L组、15μmol/L组、20μmol/L组、40μmol/L组在24 h、48 h、72 h对NCI-H1975细胞增殖抑制率明显较高(P<0.05),并且具有剂量及时间依赖性;对NCI-H1975细胞TKI耐药性的逆转倍数比较,预处理+姜黄素+吉非替尼组>姜黄素+吉非替尼组>预处理+吉非替尼组(P<0.05)。与对照组比较,各组NCI-H1975细胞凋亡率较高,其中联合用药组>吉非替尼组>姜黄素组(P<0.05)。与对照组比较,各组NCI-H1975细胞p-PI3K、p-Akt、p-Ras、p-ERK蛋白表达水平较低,其中联合用药组<吉非替尼组<姜黄素组(P<0.05)。结论姜黄素能有效逆转NCI-H1975细胞TKI靶向药物耐药性,抑制细胞增殖并促进其凋亡,最佳给药方式是姜黄素预处理后再与吉非替尼联用,作用机制可能与下调p-PI3K、p-Akt、p-Ras、p-ERK蛋白水平有关。
Objective To investigate the molecular mechanism of curcumin in reversing the drug resistance of tyrosine kinase inhibitor(TKI) targeted drugs in non-small cell lung cancer(NSCLC). Methods NCI-H1975 cells of NSCLC were selected and treated with corresponding drugs. MTT assay was used to detect the inhibitory effect of curcumin on the proliferation of NCI-H1975 cells. The reversal effect of curcumin on NCI-H1975 cell resistance was also detected. The effect of curcumin on the apoptosis rate of NCI-H1975 cells was detected by flow cytometry. The expression levels of p-PI3K, p-Akt, p-Ras and p-ERK proteins in NCI-H1975 cells were detected by Western Blot method. Results Compared with the control group, the cell proliferation inhibitory rate of NCI-H1975 cells was significantly higher in the curcumin 5 μmol/L group, 10 μmol/L group, 15 μmol/L group, 20 μmol/L group and 40 μmol/L group at 24 h, 48 h and 72 h(P<0.05), showing a dose and time dependence; the reversing fold of TKI resistance of NCI-H1975 cells was compared, and pretreatment+curcumin+gefitinib group>curcumin+gefitinib group>pretreatment+gefitinib group(P<0.05).Compared with the control group, the apoptosis rate of NCI-H1975 cells in each group was higher, and the combined drug use group>gefitinib group>curcumin group(P<0.05). Compared with the control group, the expression levels of pPI3 K, p-Akt, p-Ras and p-ERK protein in NCI-H1975 cells in each group were lower, and the combined drug use group gefitinib group<curcumin group(P<0.05). Conclusion Curcumin can effectively reverse the drug resistance of NCI-H1975 cells to TKI, inhibit cell proliferation and promote its apoptosis. The optimal administration route is the combined use with gefitinib after curcumin pretreatment. The mechanism may be related to down-regulation of p-PI3 K,p-Akt, p-Ras, p-ERK protein levels.
出处
《中国现代医生》
2017年第25期37-41,169,共6页
China Modern Doctor
基金
浙江省自然科学基金资助项目(LY13H160037)
关键词
姜黄素
非小细胞肺癌
TKI靶向药物
耐药机制
Curcumin
Non-small cell lung cancer
TKI targeted drugs
Resistance mechanism
