Bak基因转染对乳腺癌MCF-7细胞增殖及对紫杉醇敏感性的影响

邵华 冯传波 王钟林 朱双九 张英 朱明珍

解剖学报 ›› 2019, Vol. 50 ›› Issue (6) : 761-765.

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解剖学报 ›› 2019, Vol. 50 ›› Issue (6) : 761-765. DOI: 10.16098/j.issn.0529-1356.2019.06.010
肿瘤生物学

Bak基因转染对乳腺癌MCF-7细胞增殖及对紫杉醇敏感性的影响

  • 邵华1 冯传波1 王钟林1 朱双九1 张英2 朱明珍3*
作者信息 +

Effect of Bak gene overexpression on proliferation of breast cancer MCF-7 cells and sensitivity to paclitaxel

  • SHAO Hua1 FENG Chuan-bo1 WANG Zhong-lin1 ZHU Shuang-jiu1 ZHANG Ying2 ZHU Ming-zhen 3*
Author information +
文章历史 +

摘要

目的 探讨Bak基因转染对乳腺癌MCF-7细胞增殖及对紫杉醇敏感性的影响。 方法 采用Western blotting法和Real-time PCR检测乳腺癌MCF-7细胞转染前后Bak蛋白表达。通过细胞计数试剂盒8(CCK-8)和流式细胞术检测Bak基因转染后,及紫杉醇作用24 h、48 h和72 h后对MCF-7细胞增殖和细胞周期的影响。 结果 Western blotting和 Real-time PCR检测结果显示,转染质粒后,Bak蛋白表达量显著升高,MCF-7 Bak组mRNA表达量为2.15±0.07,明显高于MCF-7 NC组1.03±0.04(t=13.412,P<0.05)。转染48 h、72 h和96 h后,MCF-7 Bak细胞增殖速率为(0.31±0.03)%、(0.37±0.03)%、(0.47±0.04)%,低于MCF-7 NC组的(0.40±0.03)%、0.48±0.04)%、(0.61±0.06)%,差异有统计学意义(t 48=2.145、t 72=3.164、t 96=5.487,P<0.05)。MCF-7Bak组G2期细胞数是(26.84±2.69)%,显著高于MCF-7 NC组(16.02±1.61)%(t=12.887,P<0.05)。紫杉醇作用24 h、48 h和72 h后,MCF-7 Bak组细胞增殖抑制率为(35.98±4.00)%、(54.66±5.50)%、(80.11±8.00)%,高于MCF-7 NC组的(24.12±2.40)%、(40.12±4.00)%、(61.09±6.09)%,差异有统计学意义(t 24=8.456、t 48=10.547、t 72=13.442,P<0.05)。紫杉醇作用24 h后,MCF-7 Bak组G0/G1期细胞数(73.01±7.02)%高于MCF-7 NC组(63.84±6.68)%(P<0.05)。 结论 上调Bak基因表达可抑制乳腺癌MCF-7细胞增殖,上调G0/G1期比例,增强紫杉醇的敏感性。

Abstract

Objective To investigate the effect of Bak gene overexpression on proliferation of breast cancer MCF-7 cells and sensitivity to paclitaxel. Methods Western blotting and Real-time PCR were used to detect the expression of Bak protein in breast cancer MCF-7 cells before and after transfection. The effects of Bak gene overexpression and paclitaxel treatment for 24 hours, 48 hours and 72 hours on MCF-7 cell proliferation and cell cycle were detected by cell counting kit-8(CCK-8) and flow cytometry. Results The results of Western blotting and Real-time PCR showed that the expression of Bak protein increased significantly after transfection.The results of Western blotting and Real-time PCR showed that the expression of Bak protein increased significantly after transfection, and the mRNA expression of MCF-7-Bak group was 2.15±0.07, which was significantly higher than that of MCF-7-NC group,which was 1.03±0.04(t=13.412, P<0.05).After 48 hours, 72 hours and 96 hours of transfection, the proliferation rates of MCF-7-Bak cells were (0.31±0.03)%, (0.37±0.03)%, (0.47±0.04)%, respectively, which were lower than that of MCF-7 NC group(0.40±0.03)%, (0.48±0.04)%, (0.61±0.06)%, and the difference was statistically significant (t 48=2.145, t 72=3.164, t 96=5.487, P<0.05). The number of G2 phase cells in MCF-7 Bak group were (26.84±2.69)% significantly higher than that in MCF-7 NC group (16.02±1.61)% (t=12.887, P<0.05). After 24 hours, 48 hours, and 72 hours of paclitaxel treatment, the cell inhibition rate of MCF-7-Bak group was (35.98±4.00)%, (54.66±5.50)%, (80.11±8.00)%, respectively, which were higher than that of MCF-7 NC group (24.12±2.40)%, (40.12±4.00)%,(61.09±6.09)%. And the difference was statistically significant (t 24=8.456, t 48=10.547, t 72=13.442, P<0.05). After 24 hours of paclitaxel treatment, the number of G2 phase cells in MCF-7 Bak group was (73.01±7.02)% higher than that in MCF-7 NC group (63.84±6.68) %(t=9.736, P<0.05). Conclusion Up-regulate of Bak gene expression can inhibit the proliferation of breast cancer MCF-7 cells, up-regulating the proportion of G0/G1 phase, and enhance the sensitivity of paclitaxel.

关键词

Bak / MCF-7细胞 / 紫杉醇 / 敏感性 / 免疫印迹法 /

Key words

Bak / MCF-7 cell / Paclitaxel / Sensitivity / Western blotting / Human

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邵华 冯传波 王钟林 朱双九 张英 朱明珍. Bak基因转染对乳腺癌MCF-7细胞增殖及对紫杉醇敏感性的影响[J]. 解剖学报. 2019, 50(6): 761-765 https://doi.org/10.16098/j.issn.0529-1356.2019.06.010
SHAO Hua FENG Chuan-bo WANG Zhong-lin ZHU Shuang-jiu ZHANG Ying ZHU Ming-zhen. Effect of Bak gene overexpression on proliferation of breast cancer MCF-7 cells and sensitivity to paclitaxel[J]. Acta Anatomica Sinica. 2019, 50(6): 761-765 https://doi.org/10.16098/j.issn.0529-1356.2019.06.010

参考文献

 [1] Danova M, Chiroli S, Rosti G, et al. Cost-effectiveness of pegfilgrastim versus six days of filgrastim for preventing febrile neutropenia in breast cancer patients[J]. Tumori, 2016, 95(2):219-226.
 [2] Guo ChM, Wu N, Guo LY. Prognostic factors for HER-2-positive breast cancer in China: a Meta-analysis[J]. Chinese Journal of General Surgery, 2016, 25(2):264-270. (in Chinese)
郭晨明, 吴楠, 郭丽英. 我国HER-2阳性乳腺癌预后影响因素的Meta分析[J]. 中国普通外科杂志, 2016, 25(2):264-270.
 [3] Walker CH, Drew BA, Antoon JW, et al. Neurocognitive effects of chemotherapy and endocrine therapies in the treatment of breast cancer: recent perspectives[J]. Cancer Invest, 2016, 30(2):135.
 [4] Xu XW, Yang YL, Sun ZhL, et al. Raptor’s role in invasion and metastasis of breast cancer through Wnt3a/beta-catenin signaling pathway [J].Acta Anatomica Sinica, 2017, 48 (6): 682-687.  (in Chinese)
徐新伟,杨玉玲,孙志亮,等.Raptor通过Wnt3a/β-catenin信号通路对乳腺癌侵袭与转移的作用[J].解剖学报,2017,48(6):682-687.
 [5] Wang LN, Zhang ChJ, Li LF, et al. Clinical observation of neoadjuvant chemotherapy with paclitaxel administered weekly combined with epirubicin and cyclophosphamide regimen in triple-negative breast cancer[J]. Chinese Journal of Cancer Prevention and Treatment, 2015, 22(3):211-215.  (in Chinese)
王丽娜, 张崇建, 李连方,等. 三阴性乳腺癌表柔比星和环磷酰胺联合紫杉醇周疗新辅助化疗临床观察[J]. 中华肿瘤防治杂志, 2015, 22(3):211-215.
 [6] Wu J, Shao ZM, Shen ZZ, et al. Significance of apoptosis and apoptotic-related proteins, Bcl-2, and Bax in primary breast cancer[J]. Breast J, 2015, 6(1):44-52.
 [7] Veneti E, Tu RS, Auguste DT. RGD-targeted liposome binding and uptake on breast cancer cells is dependent on elastin linker secondary structure[J]. Bioconjug Chem, 2016, 27(8):1813-1821.
 [8] Sun JF,He HT.Mechanism of baicalein induced apoptosis in laryngeal carcinoma cells in vitro by mitochondrial apoptosis pathway[J].Chinese Journal of Gerontology,2018,38(8):1947-1951.  (in Chinese)
孙吉凤,何海涛.黄芩素通过线粒体凋亡途径诱导体外培养喉癌细胞凋亡的机制[J].中国老年学杂志,2018,38(8):1947-1951.
 [9] Jia YL,He QS, Feng Y, et al. Impacts of small Banxia plus fuling ethanol extract on mitochondrial apoptotic pathway of HepG2 and BGC823[J]. Chongqing Medicine, 2017, 46(3):296-298.  (in Chinese)
贾亚玲, 何前松, 冯泳,等. 小半夏加茯苓醇提物对HepG2、BGC823线粒体凋亡途径的影响[J]. 重庆医学, 2017, 46(3):296-298.
 [10] Li X, Zeng X, Sun J, et al. Imperatorin induces Mcl-1 degradation to cooperatively trigger Bax translocation and Bak activation to suppress drug-resistant human hepatoma[J]. Cancer Lett, 2014, 348(1-2):146-155.
 [11] Yang GX, Zhou Y, Tian H, et al. D,L-sulforaphane N-Acetyl-L-cysteine induces apoptosis by upregulating Bak and downregulating Bcl-2 in human prostate cancer cells[J]. Journal of Shanxi Medical University, 2016, 47(4):311-314. (in Chinese)
杨高祥, 周妍, 田华,等. N-乙酰半胱氨酸萝卜硫素通过上调Bak和下调Bcl-2蛋白表达诱导前列腺癌细胞凋亡[J]. 山西医科大学学报, 2016, 47(4):311-314.
 [12] Gao QL, Zhu YN, Shi B, et al.Serum containing juglans mandshurica pericarp inhibits proliferation and induces apoptosis of gastric carcinoma SGC-7901 cell[J].Chinese Journal of Experimental traditional Medical Formulae,2017,23(13):111-115. (in Chinese)
高启龙,朱亚楠,石变,等.青龙衣含药血清抑制胃癌SGC-7901细胞生长和诱导凋亡[J].中国实验方剂学杂志,2017,23(13):111-115.
 [13] Luo Y, Wang X, Wang H, et al. High bak expression is associated with a favorable prognosis in breast cancer and sensitizes breast cancer cells to paclitaxel[J]. PLoS One, 2015, 10(9):e0138955.
 [14] Zhang MW, Qin YG, Wang YQ, et al.Evaluation of efficacy and safety of bevacizumab combined with albumin-bound paclitaxel for patients with platinum-resistant or recurrent ovrain cancer[J].  Chinese Journal of Cancer Prevention and Treatment,  2016, 23(5):331-334. (in Chinese)
张孟伟, 秦亚光, 王亚秋,等. 铂类耐药复发转移性卵巢癌贝伐单抗联合白蛋白结合型紫杉醇治疗临床观察[J]. 中华肿瘤防治杂志, 2016, 23(5):331-334.
 [15] Li FH, Yang FY, Chang JY, et al. Randomized clinical study of short term efficacy and safety of paclitaxel combined with lobaplatin compared to paclitaxel combine with cisplatin in chemoradiotherapy of locally advanced cervical cancer[J]. Chinese Journal of Cancer Prevention and Treatment, 2016, 23(16):1090-1094. (in Chinese)
李凤虎, 杨飞月, 常建英,等. 宫颈癌紫杉醇联合洛铂或顺铂新辅助序贯洛铂或顺铂同步化放疗多中心随机临床研究[J]. 中华肿瘤防治杂志, 2016, 23(16):1090-1094.
 [16] Lei Y, Wan J, Li DD, et al.Down-regulation of FoxM1 increases the sensitivity of human nasopharyngeal carcinoma cells to paclitaxel by activating JNK/mitochondrial pathway[J].Journal of Third Military Medical University, 2018, 40(9): 780-787. (in Chinese)
雷越, 万婕, 李丹丹,等. 下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性[J]. 第三军医大学学报, 2018, 40(9):780-787.
 [17] Miller AV, Hicks MA, Nakajima W, et al. Paclitaxel-induced apoptosis is BAK-dependent, but BAX and BIM-independent in breast tumor[J]. PLoS One, 2013, 8(4):e60685.

基金

基金项目:江苏省连云港市科技局重点学科建设项目(SH1526)

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