Evaluation of Reversal Effects of Four C21Steroids in Marsdenia tenacissima on Paclitaxel Resistance in Ovarian Cancer Cells
KONG Qian-wen1, HU Yu-jie1, XUE Xiao-chuan2, SHI Mei-zhi1, XU Ling-yan1, ZHOU Yang-yun1, ZHANG Min1, HU Jiu-dong1, YANG Jiao1, HAN Yong-long1*, CHEN Jun-jun1*
1. Department of Pharmacy,Shanghai Jiao Tong University Affiliated Sixth People′s Hospital, Shanghai 200030, China; 2. College of Food Science and Technology, Shanghai Ocean University, Shanghai 200135, China
Abstract：OBJECTIVE To evaluate four C21 steroids in Marsdenia tenacissima, tenacissoside I (Tsd-I), tenacissoside G (Tsd-G), tenacissoside H (Tsd-H), 17β-tenacigenin B (17β-ten-B), on reversing paclitaxel resistance of the human paclitaxel-resistant ovarian cancer cells A2780/Taxol (A2780/T), and explore the underlying mechanism. METHODS Human ovarian cancer cells A2780/T were divided into four groups: control group, C21 steroid monotherapy group, PTX monotherapy group, and C21 steroid combined with PTX group (1∶1). Cell proliferation ability was detected by MTT assay. The relationship between combined action (Fa) and combination index (CI) of the two agents was analyzed by the median-effect principle. Furthermore, cell migration was observed by cell scratch experiment. Flow cytometry and Hoechst 33258 staining were used to observe the effects of drugs on cell cycle, apoptosis and apoptosis morphology. Simultaneously, the mRNA expressions of ABCB1, SLC4A2 and SLCO1A2 was detected by RT-PCR, and the P-gp level was detected by Western blot.Drug efflux assay was performed to detect the accumulation of PTX and rhodamine 123 in A2780/T cells. RESULTS C21 steroid alone or in combination with PTX could suppress the proliferation of A2780/T cells with a dose-dependent effect. Tsd-G, Tsd-H combined with PTX for 48 h had synergistic effect, and could significantly inhibit A2780/T cells migration, arrest cells in G2/M phase, promote cell apoptosis, reduce ABCB1, SLC4A2, SLCO1A2 mRNA level, and further decrease P-gp expression level which significantly promoted PTX and Rhodamine 123 accumulation in A2780/T cells. CONCLUSION The combination of Tsd-G and Tsd-H with PTX respectively can reverse the paclitaxel-resistance of human ovarian cancer A2780/T cells.The mechanism may be related to the inhibition of the expression levels of ABCB1, SLC4A2, SLCO1A2 mRNA and P-gp, and the promotion of PTX accumulation in A2780/T cells.
孔欠文, 胡玉洁, 薛晓川, 石美智, 徐玲艳, 周阳云, 张敏, 胡九东, 杨姣, 韩永龙, 陈君君. 通关藤中4种C21甾体化合物逆转卵巢癌细胞紫杉醇耐药的作用评价[J]. 中国药学杂志, 2022, 57(9): 706-717.
KONG Qian-wen, HU Yu-jie, XUE Xiao-chuan, SHI Mei-zhi, XU Ling-yan, ZHOU Yang-yun, ZHANG Min, HU Jiu-dong, YANG Jiao, HAN Yong-long, CHEN Jun-jun. Evaluation of Reversal Effects of Four C21Steroids in Marsdenia tenacissima on Paclitaxel Resistance in Ovarian Cancer Cells. Chinese Pharmaceutical Journal, 2022, 57(9): 706-717.
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