吴茱萸碱、小檗碱对SGC-7901/DDP耐药性及耐药相关蛋白的影响
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摘要
目的:探讨吴茱萸碱、小檗碱及其联合使用对顺铂(Cisplatin, DDP)耐药胃癌细胞SGC7901/DDP敏感性的作用。方法:体外诱导建立人胃癌顺铂耐药细胞株SGC-7901/DDP,实验分为对照组、DDP组、吴茱萸碱+DDP组、小檗碱+DDP组及吴茱萸碱+小檗碱+DDP组,CCK8检测各组SGC-7901/DDP细胞增殖能力;Western Blot分析耐药及凋亡相关蛋白P-gp、MRP、caspase-3、caspase-9的表达。结果:CCK8实验表明吴茱萸碱、小檗碱在不影响SGC-7901/DDP细胞活力的情况下可显著增强DDP抑制细胞增殖的作用,且小檗碱化疗敏感性的作用明显强于二者单独给药(P <0.001);Western Blot结果表明,SGC-7901/DDP细胞经吴茱萸碱、小檗碱处理后,caspase-3、caspase-9表达上调,P-gp、MRP表达下调。结论:吴茱萸碱、小檗碱单用及其联用可在体外显著增强SGC-7901/DDP细胞对DDP的敏感性,其效应与凋亡蛋白caspase-3、caspase-9表达升高,耐药相关蛋白P-gp、MRP表达下调相关。
Objective: To investigate the effects of evodiamine, berberine and its combination on the sensitivity of Cisplatin(DDP)-resistant gastric cancer cell SGC7901/DDP to DDP. Methods: Human gastric cancer cisplatin-resistant cell SGC-7901/DDP was induced in vitro and divided into control group, DDP group, evodiamine + DDP group,berberine + DDP group and evodiamine + berberine + DDP group. CCK8 was used to detect the proliferation of SGC-7901/DDP cells. Western Blot was used to analyze the expression of drug-resistant and apoptosis-related proteins including P-gp, MRP, caspase-3 and caspase-9. Results: Evodiamine or berberine combined with DDP could significantly enhance the inhibition of DDP on cell proliferation without affecting the viability of SGC-7901/DDP cells,combination treatment of berberine(8μM) and evodiamine(3.2 μM) significantly increased chemosensitivity of SGC-7901/DDP than that of either alone(P < 0.001). The results of Western Blot showed that evodiamine or berberine upregulated the expression of caspase-3 and caspase-9, while the expression of drug-related proteins P-gp and MRP was down-regulated. Conclusion: The combination of evodiamine, berberine and DDP can significantly enhance the sensitivity of SGC-7901/DDP cells to DDP in vitro, which was associated with increased expression of caspase-3 and caspase-9, and down-regulation of resistance-related proteins P-gp and MRP.
Objective: To investigate the effects of evodiamine, berberine and its combination on the sensitivity of Cisplatin(DDP)-resistant gastric cancer cell SGC7901/DDP to DDP. Methods: Human gastric cancer cisplatin-resistant cell SGC-7901/DDP was induced in vitro and divided into control group, DDP group, evodiamine + DDP group,berberine + DDP group and evodiamine + berberine + DDP group. CCK8 was used to detect the proliferation of SGC-7901/DDP cells. Western Blot was used to analyze the expression of drug-resistant and apoptosis-related proteins including P-gp, MRP, caspase-3 and caspase-9. Results: Evodiamine or berberine combined with DDP could significantly enhance the inhibition of DDP on cell proliferation without affecting the viability of SGC-7901/DDP cells,combination treatment of berberine(8μM) and evodiamine(3.2 μM) significantly increased chemosensitivity of SGC-7901/DDP than that of either alone(P < 0.001). The results of Western Blot showed that evodiamine or berberine upregulated the expression of caspase-3 and caspase-9, while the expression of drug-related proteins P-gp and MRP was down-regulated. Conclusion: The combination of evodiamine, berberine and DDP can significantly enhance the sensitivity of SGC-7901/DDP cells to DDP in vitro, which was associated with increased expression of caspase-3 and caspase-9, and down-regulation of resistance-related proteins P-gp and MRP.
引文
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3 Sarkar F H, Li Y. Harnessing the fruits of nature for the development of multi-targeted cancer therapeutics. Cancer Treat Rev. 2009, 35:597–607.
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5 吴亚琼,方伟蓉,李运曼.肿瘤多药耐药机制及逆转药物的研究进展.药学与临床研究, 2016, 1:43-47.
6 王冰,宋锐,牟宗玲,等.中药逆转肿瘤多药耐药的相关研究.中医学报, 2016, 31(7):946-948.
7 Shan X, Aziz F, Tian L L, et al. Ginsenoside Rg3-induced EGFR/MAPK pathway deactivation inhibits melanoma cell proliferation by decreasing FUT4/LeY expression. Int J Oncol, 2015, 46:1667-1676.
8 Shan X, Fu Y S, Aziz F, et al. Ginsenoside Rg3 inhibits melanoma cell proliferation through down-regulation of histone deacetylase 3(HDAC3)and increase of p53 acetylation. PLoS One, 2014, 9:e115401.
9 Lee Y J, Lee S, Ho J N, et al. Synergistic antitumor effect of ginsenoside Rg3 and cisplatin in cisplatin resistant bladder tumor cell line. Oncol Rep, 2014, 32:1803-1808.
10 Zhang Q, Kang X, Zhao W. Antiangiogenic effect of low-dose cyclophosphamide combined with ginsenoside Rg3 on Lewis lung carcinoma. Biochem Biophys Res Commun, 2006, 342:824-828.
11 Sui H, Duan P, Guo P, et al. Zhi Zhen Fang formula reverses Hedgehog pathway mediated multidrug resistance in colorectal cancer. Oncol Rep,2017, 38(4):2087-2095.
12 周祥羽,邹忠杰.左金丸化学成分及现代药理研究进展.广东化工,2017, 44(10):89-90.
13 Xu L, Qi Y, Lv L, et al. In vitro anti-proliferative effects of Zuojinwan on eight kinds of human cancer cell lines. Cytotechnology, 2014, 66(1):37-50.
14 Du J, Sun Y, Wang X F, et al. Establishment of an experimental breast cancer ZHENG model and curative effect evaluation of Zuo-Jin Wan.Evid Based Complement Alternat Med, 2013, 2013:324732.
15 Tang Q F, Sun J, Yu H, et al. The Zuo Jin Wan formula induces mitochondrial apoptosis of cisplatin-resistant gastric cancer cells via Cofilin-1. Evid Based Complement Alternat Med, 2016, 2016:8203789.
16 Hesari A, Ghasemi F, Cicero A F G, et al. Berberine:A potential adjunct for the treatment of gastrointestinal cancers? J Cell Biochem,2018, 119(12):9655-9663.
17 Wang Y, Zhang S. Berberine suppresses growth and metastasis of endometrial cancer cells via miR-101/COX-2. Biomed Pharmacother,2018, 103:1287-1293.
18 Zhang T, Qu S, Shi Q, et al. Evodiamine induces apoptosis and enhances TRAIL-induced apoptosis in human bladder cancer cells through mTOR/S6K1-mediated downregulation of Mcl-1. Int J Mol Sci,2014, 15(2):3154-3171.
19 Shi H L, Wu X J, Liu Y, et al. Berberine counteracts enhanced IL-8expression of AGS cells induced by evodiamine. Life Sci, 2013, 93(22):830-839.
20 Zhang R, Qiao H, Chen S, et al. Berberine reverses lapatinib resistance of HER2-positive breast cancer cells by increasing the level of ROS.Cancer Biol Ther, 2016, 17(9):925-934.
21 Hu C, Gao X, Han Y, et al. Evodiamine sensitizes BGC-823 gastric cancer cells to radiotherapy in vitro and in vivo. Mol Med Rep, 2016, 14(1):413-419.
22 Yang L, Liu X, Wu D, et al. Growth inhibition and induction of apoptosis in SGC7901 human gastric cancer cells by evodiamine. Mol Med Rep, 2014, 9(4):1147-1152.
23 Hu C Y, Wu H T, Su Y C, et al. Evodiamine exerts an antihepatocellular carcinoma activity through a WWOX-dependent pathway. Molecules, 2017, 22(7):1-8.
24 Du J, Wang X F, Zhou Q M, et al. Evodiamine induces apoptosis and inhibits metastasis in MDA-MB-231 human breast cancer cells in vitro and in vivo. Oncol Rep, 2013, 30(2):685-694.
25 Zhong Z F, Tan W, Wang S P, et al. Anti-proliferative activity and cell cycle arrest induced by evodiamine on paclitaxel-sensitive andresistant human ovarian cancer cells. Sci Rep, 2015, 5:16415.
26 Sui H, Zhou L H, Zhang Y L, et al. Evodiamine suppresses ABCG2mediated drug resistance by inhibiting p50/p65 NF-κB pathway in colorectal cancer. J Cell Biochem, 2016, 117(6):1471-1481.
27 Liu S, Fang Y, Shen H, et al. Berberine sensitizes ovarian cancer cells to cisplatin through miR-21/PDCD4 axis. Acta Biochim Biophys Sin(Shanghai), 2013, 45:756-762.
28 李钧.小檗碱抗肿瘤研究进展.科学技术创新, 2017(14):119-120.
29 Yu M, Tong X, Qi B, et al. Berberine enhances chemosensitivity to irinotecan in colon cancer via inhibition of NF-κB. Mol Med Rep, 2014,9(1):249-254.
30 Liang K W, Yin S C, Ting CT, et al. Berberine inhibits platelet-derived growth factor-induced growth and migration partly through an AMPKdependent pathway in vascular smooth muscle cells. Eur J Pharmacol,2008, 590(1-3):343-354.
31 Chen Q, Qin R, Fang Y, et al. Berberine sensitizes human ovarian cancer cells to Cisplatin through miR-93/PTEN/Akt signaling pathway.Cell Physiol Biochem, 2015, 36(3):956-965.
32 Wang M, Liu Z F, Tang H, et al. Application of alkaloids in reversing multidrug resistance in human cancers. Chinese Journal of Natural Medicines, 2018, 16(8):561-571.
33 You H Y, Xie X M, Zhang W J, et al. Berberine modulates cisplatin sensitivity of human gastric cancer cells by upregulation of miR-203. In Vitro Cell Dev Biol Anim, 2016, 52(8):857-863.
35 徐俊杰,杨然,杨芳景,等.吴茱萸碱抗肿瘤机制的研究进展.上海交通大学学报(医学版), 2018, 38(5):578-583.
35 Velingkar V S, Dandekar V D. Modulation of P-Glyco protein Mediated Multidrug Resistance(Mdr)in Cancer Using Chemosensitizers. IJPSR,2010, 1(2):104-111.
36 Glavinas H, Krajcsi P, Cserepes J, et al. The role of ABC transporters in drugresistance,metabolismandtoxicity. Curr Drug Deliv, 2004,1(1):27-42.
37 Xu H W, Xu L, Hao J H, et al. Expression of P-glycoprotein and multidrug resistance-associated protein is associated with multidrug resistance in gastric cancer. J Int Med Res, 2010, 38(1):34-42.
2 Venerito M, Link A, Rokkas T, et al. Gastric cancer-clinical and epidemiological aspects. Helicobacter, 2016, 1:39-44.
3 Sarkar F H, Li Y. Harnessing the fruits of nature for the development of multi-targeted cancer therapeutics. Cancer Treat Rev. 2009, 35:597–607.
4 Wu Q, Yang Z, Nie Y, et al. Multi-drug resistance in cancer chemotherapeutics:mechanisms and lab approaches. Cancer Lett, 2014,347(2):159-166.
5 吴亚琼,方伟蓉,李运曼.肿瘤多药耐药机制及逆转药物的研究进展.药学与临床研究, 2016, 1:43-47.
6 王冰,宋锐,牟宗玲,等.中药逆转肿瘤多药耐药的相关研究.中医学报, 2016, 31(7):946-948.
7 Shan X, Aziz F, Tian L L, et al. Ginsenoside Rg3-induced EGFR/MAPK pathway deactivation inhibits melanoma cell proliferation by decreasing FUT4/LeY expression. Int J Oncol, 2015, 46:1667-1676.
8 Shan X, Fu Y S, Aziz F, et al. Ginsenoside Rg3 inhibits melanoma cell proliferation through down-regulation of histone deacetylase 3(HDAC3)and increase of p53 acetylation. PLoS One, 2014, 9:e115401.
9 Lee Y J, Lee S, Ho J N, et al. Synergistic antitumor effect of ginsenoside Rg3 and cisplatin in cisplatin resistant bladder tumor cell line. Oncol Rep, 2014, 32:1803-1808.
10 Zhang Q, Kang X, Zhao W. Antiangiogenic effect of low-dose cyclophosphamide combined with ginsenoside Rg3 on Lewis lung carcinoma. Biochem Biophys Res Commun, 2006, 342:824-828.
11 Sui H, Duan P, Guo P, et al. Zhi Zhen Fang formula reverses Hedgehog pathway mediated multidrug resistance in colorectal cancer. Oncol Rep,2017, 38(4):2087-2095.
12 周祥羽,邹忠杰.左金丸化学成分及现代药理研究进展.广东化工,2017, 44(10):89-90.
13 Xu L, Qi Y, Lv L, et al. In vitro anti-proliferative effects of Zuojinwan on eight kinds of human cancer cell lines. Cytotechnology, 2014, 66(1):37-50.
14 Du J, Sun Y, Wang X F, et al. Establishment of an experimental breast cancer ZHENG model and curative effect evaluation of Zuo-Jin Wan.Evid Based Complement Alternat Med, 2013, 2013:324732.
15 Tang Q F, Sun J, Yu H, et al. The Zuo Jin Wan formula induces mitochondrial apoptosis of cisplatin-resistant gastric cancer cells via Cofilin-1. Evid Based Complement Alternat Med, 2016, 2016:8203789.
16 Hesari A, Ghasemi F, Cicero A F G, et al. Berberine:A potential adjunct for the treatment of gastrointestinal cancers? J Cell Biochem,2018, 119(12):9655-9663.
17 Wang Y, Zhang S. Berberine suppresses growth and metastasis of endometrial cancer cells via miR-101/COX-2. Biomed Pharmacother,2018, 103:1287-1293.
18 Zhang T, Qu S, Shi Q, et al. Evodiamine induces apoptosis and enhances TRAIL-induced apoptosis in human bladder cancer cells through mTOR/S6K1-mediated downregulation of Mcl-1. Int J Mol Sci,2014, 15(2):3154-3171.
19 Shi H L, Wu X J, Liu Y, et al. Berberine counteracts enhanced IL-8expression of AGS cells induced by evodiamine. Life Sci, 2013, 93(22):830-839.
20 Zhang R, Qiao H, Chen S, et al. Berberine reverses lapatinib resistance of HER2-positive breast cancer cells by increasing the level of ROS.Cancer Biol Ther, 2016, 17(9):925-934.
21 Hu C, Gao X, Han Y, et al. Evodiamine sensitizes BGC-823 gastric cancer cells to radiotherapy in vitro and in vivo. Mol Med Rep, 2016, 14(1):413-419.
22 Yang L, Liu X, Wu D, et al. Growth inhibition and induction of apoptosis in SGC7901 human gastric cancer cells by evodiamine. Mol Med Rep, 2014, 9(4):1147-1152.
23 Hu C Y, Wu H T, Su Y C, et al. Evodiamine exerts an antihepatocellular carcinoma activity through a WWOX-dependent pathway. Molecules, 2017, 22(7):1-8.
24 Du J, Wang X F, Zhou Q M, et al. Evodiamine induces apoptosis and inhibits metastasis in MDA-MB-231 human breast cancer cells in vitro and in vivo. Oncol Rep, 2013, 30(2):685-694.
25 Zhong Z F, Tan W, Wang S P, et al. Anti-proliferative activity and cell cycle arrest induced by evodiamine on paclitaxel-sensitive andresistant human ovarian cancer cells. Sci Rep, 2015, 5:16415.
26 Sui H, Zhou L H, Zhang Y L, et al. Evodiamine suppresses ABCG2mediated drug resistance by inhibiting p50/p65 NF-κB pathway in colorectal cancer. J Cell Biochem, 2016, 117(6):1471-1481.
27 Liu S, Fang Y, Shen H, et al. Berberine sensitizes ovarian cancer cells to cisplatin through miR-21/PDCD4 axis. Acta Biochim Biophys Sin(Shanghai), 2013, 45:756-762.
28 李钧.小檗碱抗肿瘤研究进展.科学技术创新, 2017(14):119-120.
29 Yu M, Tong X, Qi B, et al. Berberine enhances chemosensitivity to irinotecan in colon cancer via inhibition of NF-κB. Mol Med Rep, 2014,9(1):249-254.
30 Liang K W, Yin S C, Ting CT, et al. Berberine inhibits platelet-derived growth factor-induced growth and migration partly through an AMPKdependent pathway in vascular smooth muscle cells. Eur J Pharmacol,2008, 590(1-3):343-354.
31 Chen Q, Qin R, Fang Y, et al. Berberine sensitizes human ovarian cancer cells to Cisplatin through miR-93/PTEN/Akt signaling pathway.Cell Physiol Biochem, 2015, 36(3):956-965.
32 Wang M, Liu Z F, Tang H, et al. Application of alkaloids in reversing multidrug resistance in human cancers. Chinese Journal of Natural Medicines, 2018, 16(8):561-571.
33 You H Y, Xie X M, Zhang W J, et al. Berberine modulates cisplatin sensitivity of human gastric cancer cells by upregulation of miR-203. In Vitro Cell Dev Biol Anim, 2016, 52(8):857-863.
35 徐俊杰,杨然,杨芳景,等.吴茱萸碱抗肿瘤机制的研究进展.上海交通大学学报(医学版), 2018, 38(5):578-583.
35 Velingkar V S, Dandekar V D. Modulation of P-Glyco protein Mediated Multidrug Resistance(Mdr)in Cancer Using Chemosensitizers. IJPSR,2010, 1(2):104-111.
36 Glavinas H, Krajcsi P, Cserepes J, et al. The role of ABC transporters in drugresistance,metabolismandtoxicity. Curr Drug Deliv, 2004,1(1):27-42.
37 Xu H W, Xu L, Hao J H, et al. Expression of P-glycoprotein and multidrug resistance-associated protein is associated with multidrug resistance in gastric cancer. J Int Med Res, 2010, 38(1):34-42.