丹参酮ⅡA对人结肠癌HT29细胞TGF-β1、Bcl-2表达和细胞生长的影响
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摘要
目的观察丹参酮ⅡA对人结肠癌细胞HT29生长及其对转化生长因子β1(TGF-β1)、B细胞淋巴瘤-2(Bcl-2)基因表达的影响,为结肠癌治疗提供理论基础。方法分别采用不同浓度的丹参酮ⅡA处理人结肠癌HT29细胞,MTT法检测细胞生长情况;实时荧光定量PCR(Real-time PCR)检测不同处理组所收集细胞TGF-β1和Bcl-2 mRNA表达;小鼠腋下接种HT29细胞制备小鼠结肠癌移植瘤模型,随机分为对照组和丹参酮ⅡA处理组,每周测量肿瘤体积1次,4周后摘取肿瘤称重并保存肿瘤;Western blot检测提取自HT29细胞和肿瘤组织内TGF-β1、Bcl-2蛋白表达。结果随着丹参酮ⅡA浓度升高,HT29生长抑制率增加;丹参酮ⅡA浓度增加对TGF-β1和Bcl-2 mRNA及蛋白表达产生抑制作用;丹参酮ⅡA对小鼠结肠癌移植瘤的体积抑制率为28.63%;Western blot结果表明,与对照组比较,丹参酮ⅡA组肿瘤体积内TGF-β1、Bcl-2蛋白水平降低。结论丹参酮ⅡA对结肠癌细胞HT29及其裸鼠模型移植瘤的生长具有明显的抑制作用,其机制可能与丹参酮ⅡA下调TGF-β1和Bcl-2表达有关。
Objective To observe the effects of Tanshinone ⅡA on the growth of HT29 and the expressions of TGF-β1 and Bcl-2 in colon cancer so as to provide theoretical assistance for treatment of colon cancer. Methods Human colon cancer HT29 cells were cultured for different time after treated with various concentrations of Tanshinone ⅡA, respectively. MTT method was used to detect cell growth. The human colon cancer tumors on nude mice were randomly divided into control group and Tanshinone ⅡA treatment group, and the tumor volume was measured once a week. After 4 weeks, the tumors were preserved. The levels of TGF-β1 and Bcl-2 protein in the HT29 cells and tumor tissues were detected by Western blot. Results With the increase of Tanshinone ⅡA concentration, the growth inhibition rate of HT29 increased; Tanshinone ⅡA downregulated the mRNA and protein levels of TGF-β1 and Bcl-2 gene in HT29. The volume inhibition rate of Tanshinone ⅡA on mouse colon cancer transplanted tumor was 28.63%. The results of Western blot showed that the levels of TGF-β1 and Bcl-2 protein were decreased in Tanshinone ⅡA group compared with control group. Conclusion Tanshinone ⅡA has significant inhibitory effects on the growth of colon cancer cell HT29 and its xenografts model. This mechanism may be realized by the downregulation of TGF-β1 and Bcl-2 expressions via Tanshinone ⅡA.
Objective To observe the effects of Tanshinone ⅡA on the growth of HT29 and the expressions of TGF-β1 and Bcl-2 in colon cancer so as to provide theoretical assistance for treatment of colon cancer. Methods Human colon cancer HT29 cells were cultured for different time after treated with various concentrations of Tanshinone ⅡA, respectively. MTT method was used to detect cell growth. The human colon cancer tumors on nude mice were randomly divided into control group and Tanshinone ⅡA treatment group, and the tumor volume was measured once a week. After 4 weeks, the tumors were preserved. The levels of TGF-β1 and Bcl-2 protein in the HT29 cells and tumor tissues were detected by Western blot. Results With the increase of Tanshinone ⅡA concentration, the growth inhibition rate of HT29 increased; Tanshinone ⅡA downregulated the mRNA and protein levels of TGF-β1 and Bcl-2 gene in HT29. The volume inhibition rate of Tanshinone ⅡA on mouse colon cancer transplanted tumor was 28.63%. The results of Western blot showed that the levels of TGF-β1 and Bcl-2 protein were decreased in Tanshinone ⅡA group compared with control group. Conclusion Tanshinone ⅡA has significant inhibitory effects on the growth of colon cancer cell HT29 and its xenografts model. This mechanism may be realized by the downregulation of TGF-β1 and Bcl-2 expressions via Tanshinone ⅡA.
引文
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[2] 王江峰,陈晟.丹参酮ⅡA诱导人小细胞肺癌细胞凋亡及其分子机制[J].医学研究杂志,2017,46(10):84-87.
[3] 杜睿,郑鸿,王艳萍,等.丹参酮ⅡA诱导白血病NB4细胞分化分子机制研究[J].中国中药杂志,2008,33(24):2954-2958.
[4] 陈景,黄曙方,肖定璋,等.丹参酮ⅡA磺酸钠对胰腺癌BX-PC-3细胞系增殖与细胞周期调控因子表达的影响[J].中国病理生理杂志,2013,29(1):112-115.
[5] 刘丽璇,吴灵飞,邓巍,等.丹参酮ⅡA对低氧条件下人肝癌HepG2细胞增殖、凋亡的影响及与HIF-1α、VEGF和野生型P53蛋白表达的关系[J].中国病理生理杂志,2014,30(12):2155-2160.
[6] 潘登,陈可和.丹参酮ⅡA逆转多药耐药的体外效应研究[J].中国药房,2015,26(25):3488-3489,3490.
[7] 崔明,陈东来,刘茂兴,等.MSKCC结肠癌生存预测模型单中心验证性研究[J].中国实用外科杂志,2016,36(2):210-213.
[8] 伍雯,秦环龙.膳食结构改变与结肠癌风险相关性的研究进展[J].肠外与肠内营养,2014,21(1):55-59.
[9] 周利红,刘宣,王炎,等.丹参酮ⅡA对小鼠肠癌皮下移植瘤血管新生的抑制作用[J].世界华人消化杂志,2009,17(31):3203-3209.
[10] 张欣,张蒲蓉,陈洁,等.丹参酮ⅡA对乳腺癌抑制作用的体内实验研究[J].四川大学学报(医学版),2010,40(1):62-67.
[11] HARALDSDOTTIR S,EINARSDOTTIR HM,SMARADOTTIR A,et al.Colorectal cancer-review][J].Laeknabladid,2014,100(2):75-82.
[12] NAGAPPAN A,LEE WS,YUN JW,et al.Tetraarsenic hexoxide induces G2/M arrest,apoptosis,and autophagy via PI3K/Akt suppression and p38 MAPK activation in SW620 human colon cancer cells[J].PLoS One,2017,12(3):e0174591.
[13] 裴雷杰,李敬东,赵中华,等.雷公藤甲素对人口腔鳞状癌细胞增殖及PTEN基因表达的影响[J].中华口腔医学杂志,2017,52(1):44-47.
[14] 白岩.槐耳清膏抑制结肠癌切除后肿瘤复发机制探讨[J].山东医药,2013,53(38):33-35.
[15] 赵雪峰,贾楠,李勇等.丹参酮ⅡA对胃癌细胞迁移和侵袭能力的抑制作用及机制[J].中国癌症杂志,2013,(10):793-797.
[16] 李小静,李志锋,李显平,等.丹参酮ⅡA体外促进黑素瘤A375细胞自噬及信号通路的实验研究[J].中华皮肤科杂志,2017,50(1):29-32.
[17] BAI Y,ZHANG L,FANG X,et al.Tanshinone IIA enhances chemosensitivity of colon cancer cells by suppressing nuclear factor-κB[J].Exp Ther Med,2016,11(3):1085-1089.
[18] PARK SJ,LEE SK,LIM CR,et al.Heme oxygenase-1/carbon monoxide axis suppresses transforming growth factor-β1-induced growth inhibition by increasing ERK1/2-mediated phosphorylation of Smad3 at Thr-179 in human hepatocellular carcinoma cell lines[J].Biochem Biophys Res Commun,2018,498(3):609-615.
[19] WANG Y,DU C,ZHANG N,et al.TGF-β1 mediates the effects of aspirin on colonic tumor cell proliferation and apoptosis[J].Oncol Lett,2018,15(4):5903-5909.
[20] DENG B,SU F,XIE R,et al.miR-371-5p suppresses the proliferative and migratory capacity of human nasopharyngeal carcinoma by targeting BCL2[J].Oncol Lett,2018,15(6):9209-9215.
[21] LI C,PENG W,SONG X,et al.Anticancer effect of icaritin inhibits cell growth of colon cancer through reactive oxygen species,Bcl-2 and cyclin D1/E signaling[J].Oncol Lett,2016,12(5):3537-3542.