消癌平注射液联合紫杉醇对乳腺癌MCF-7细胞的增殖抑制作用及机制研究
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摘要
目的:探讨消癌平注射液联合紫杉醇对乳腺癌MCF-7细胞增殖的抑制作用及机制。方法:体外培养人乳腺癌MCF-7细胞,以不同浓度的消癌平注射液联合紫杉醇进行干预,并与消癌平注射液、紫杉醇单药组进行比较,采用MTT法、细胞克隆实验和LDH检测细胞增殖和细胞毒性,DCFH-DA荧光探针检测细胞内ROS的产生,RT-PCR检测Mfn2 mRNA的表达。结果:消癌平注射液可显著抑制MCF-7细胞的增殖和克隆,增加LDH的释放,且消癌平注射液联合紫杉醇组较紫杉醇单药组作用更为显著;消癌平注射液可显著增加ROS的生成,但联合用药组与单药组相比无统计学差异;消癌平注射液可上调Mfn2 mRNA的表达,并显著提高紫杉醇引起的Mfn2 mRNA低表达。结论:消癌平注射液联合紫杉醇可能会通过调控Mfn2 mRNA的表达,引起线粒体功能损伤并抑制乳腺癌细胞的增殖,对紫杉醇抗乳腺癌发挥增效作用。
Objective:To explore the effect and mechanism of Xiaoaiping Injection combined paclitaxel on breast cancer cells. Methods:MCF-7 cells were incubated with different concentrations of Xiaoaiping Injection combined paclitaxel and compared with single drug of Xiaoaiping Injection or paclitaxel. MTT assay, cell cloning and LDH assay were performed to detect cell proliferation and drug toxicity. DCFH-DA fluorescent probe was used to detect the intracellular ROS production. Mfn2 mRNA expression was detected by RT-PCR. Results: The data showed that Xiaoaiping Injection inhibited the proliferation and cloning of MCF-7 and increased the LDH release and ROS generation. These changes were more significantly when Xiaoaiping Injection combined paclitaxel compared with the single drug respectively except ROS. Xiaoping Injection increased the Mfn2 mRNA expression. Moreover, the decreased expression of mRNA induced by PTX could be upregulated by Xiaoaiping Injection. Conclusion:Xiaoaiping Injection combined paclitaxel could inhibit the growth of breast cancer cells and synergistic the effect of paclitaxel against breast cancer through Mfn2 mRNA induced mitochondria dysfunction.
Objective:To explore the effect and mechanism of Xiaoaiping Injection combined paclitaxel on breast cancer cells. Methods:MCF-7 cells were incubated with different concentrations of Xiaoaiping Injection combined paclitaxel and compared with single drug of Xiaoaiping Injection or paclitaxel. MTT assay, cell cloning and LDH assay were performed to detect cell proliferation and drug toxicity. DCFH-DA fluorescent probe was used to detect the intracellular ROS production. Mfn2 mRNA expression was detected by RT-PCR. Results: The data showed that Xiaoaiping Injection inhibited the proliferation and cloning of MCF-7 and increased the LDH release and ROS generation. These changes were more significantly when Xiaoaiping Injection combined paclitaxel compared with the single drug respectively except ROS. Xiaoping Injection increased the Mfn2 mRNA expression. Moreover, the decreased expression of mRNA induced by PTX could be upregulated by Xiaoaiping Injection. Conclusion:Xiaoaiping Injection combined paclitaxel could inhibit the growth of breast cancer cells and synergistic the effect of paclitaxel against breast cancer through Mfn2 mRNA induced mitochondria dysfunction.
引文
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[2] Symmans WF, Wei C, Gould R, et al. Long-Term Prognostic Risk After Neoadjuvant Chemotherapy Associated With Residual Cancer Burden and Breast Cancer Subtype[J]. Journal of clinical oncology: official journal of the American Society of Clinical Oncology, 2017, 35(10):1049-1060.
[3] 林洪生, 李萍萍, 薛冬, 等. 肿瘤姑息治疗中成药使用专家共识(2013版)[J]. 中国中西医结合杂志, 2016(3): 269-279.
[4] Coley HM. Mechanisms and strategies to overcome chemotherapy resistance in metastatic breast cancer[J]. Cancer Treatment Reviews, 2008, 34(4):378-390.
[5] Samanta D, Gilkes DM, Chaturvedi P, et al. Hypoxia-inducible factors are required for chemotherapy resistance of breast cancer stem cells[J]. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(50): E5429-5438.
[6] Bhat TA, Kumar S, Chaudhary AK, et al. Restoration of mitochondria function as a target for cancer therapy[J]. Drug Discovery Today, 2015, 20(5):635-643.
[7] Vyas S, Zaganjor E. Mitochondria and Cancer[J]. Cell, 2016, 166(3): 555-566.
[8] De Brito OM, Scorrano L. Mitofusin 2 tethers endoplasmic reticulum to mitochondria[J]. Nature, 2008, 456(7222): 605-610.
[9] Xu K, Chen G, Li X, et al. MFN2 suppresses cancer progression through inhibition of mTORC2/Akt signaling[J]. Scientific reports, 2017, 7: 41718.
[10] 郑爱文, 李涛, 陈雅卿, 等. 消癌平联合顺铂对高转移人卵巢癌 HO-8910PM 细胞的抑制作用[J]. 中华肿瘤杂志, 2016, 38(1):11-16.
[11] Han SY, Zhao MB, Zhuang GB. Marsdenia tenacissima extract restored gefitinib sensitivity in resistant non-small cell lung cancer cells[J]. Lung cancer (Amsterdam, Netherlands), 2012, 75(1):30-37.
[12] Huang Z, Wang Y, Chen J, et al.Effect of Xiaoaiping injection on advanced hepatocellular carcinoma in patients[J]. Journal of Traditional Chinese Medicine, 2013, 33(1): 34-38.
[13] Li T, Mello-Thoms C, Brennan PC. Descriptive epidemiology of breast cancer in China: incidence, mortality, survival and prevalence[J]. Breast Cancer Research and Treatment, 2016, 159(3): 395-406.
[14] 中国抗癌协会乳腺癌专业委员会.中国抗癌协会乳腺癌诊治指南与规范[S].中国癌症杂志, 2015, 25(9):692-754.
[15] Chung V CH, Wu X, Hui EP, et al. Effectiveness of Chinese herbal medicine for cancer palliative care: overview of systematic reviews with meta-analyses[J]. Scientific reports, 2015, 5:18111.
[16] 李迩娜, 王芳, 钦松, 等. 消癌平注射液抗肿瘤作用机制研究进展[J]. 中国中医药信息杂志, 2012(9): 111-112.
[17] Huang Z, Lin H, Wang Y, et al. Studies on the anti-angiogenic effect of Marsdenia tenacissima extract in vitro and in vivo[J]. Oncology Letters, 2013, 5(3):917-922.
[18] Han SY, Zhao W, Sun H, et al. Marsdenia tenacissima extract enhances gefitinib efficacy in non-small cell lung cancer xenografts[J]. Phytomedicine International Journal of Phytotherapy & Phytopharmacology, 2015, 22(5):560-567.
[19] Yang Y,Karakhanova S, Hartwig W, et al. Mitochondria and mitochondrial ROS in cancer: novel targets for anticancer therapy[J]. Journal of Cellular Physiology, 2016, 231(12):2570-2581.
[20] Takasaki T,Hagihara K,Satoh R, et al. More than Just an Immunosuppressant: The Emerging Role of FTY720 as a Novel Inducer of ROS and Apoptosis[J]. Oxidative Medicine and CellularLongevity,2018, 2018:4397159.
[21] Filadi R, Pendin D, Pizzo P. Mitofusin 2: from functions to disease[J]. Cell Death and Disease, 2018(9):330.
[22] Chen KH,Dasgupta A, Ding J, et al. Role of mitofusin 2 (Mfn2) in controlling cellular proliferation[J]. FASEB journal: official publication of the Federation of American Societies for Experimental Biology, 2014, 28(1):382-394.