四君子汤含药血清对RNA干扰后的大肠癌SW480细胞超微结构、端粒长度及h TERT表达的影响
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摘要
目的:运用四君子汤含药血清干预经RNA干扰后的大肠癌SW480细胞,观察其超微结构、端粒相对长度、端粒酶逆转录酶(hTERT)mRNA及蛋白表达变化,探讨四君子汤含药血清防治溃疡性结肠炎癌变的可能机制。方法:培养人大肠癌SW480细胞,经RNA干扰后,予低、中、高不同剂量的四君子汤含药血清,空白胎牛血清,美沙拉嗪含药血清,分别干预48h,运用透射电镜观察细胞超微结构,荧光定量PCR(SYBR Green法)和Westernblotting技术检测端粒相对长度、hTERT mRNA及蛋白表达变化。结果:经不同剂量四君子汤含药血清和美沙拉嗪含药血清干预后,RNA干扰后的大肠癌SW480细胞出现了自噬现象。各给药组端粒长度均明显小于空白组(P<0.05),其中四君子汤低剂量组端粒长度与美沙拉嗪组相当(P>0.05),中、高剂量组端粒长度大于美沙拉嗪组(P<0.05)。各给药组hTERT mRNA和蛋白表达明显低于空白组(P<0.05),其中四君子汤中剂量组hTERT mRNA表达明显低于美沙拉嗪组(P<0.05),低、高剂量组hTERT mRNA明显高于美沙拉嗪组(P<0.05),低、中剂量组hTRET蛋白表达明显低于美沙拉嗪组(P<0.05),高剂量组hTRET蛋白表达与美沙拉嗪组相当(P>0.05)。结论:四君子汤含药血清可能通过诱导细胞自噬,减缓炎症反应,下调hTERT表达,降低端粒酶活性,缩短端粒长度,抑制癌细胞增殖,达到防治溃疡性结肠炎相关癌变的效用。
引文
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[2]SMITH L A,BARAZA W,TIFFIN N,et al.Endoscopic resection of adenoma-like mass in chronic ulcerative colitis using a combined endoscopic mucosal resection and cap assisted submucosal dissection technique[J].Inflammatory Bowel Diseases,2008,14(10):1380.
[3]WOLF J M,RYBICKI L A,LASHNER B A.The impact of ursodeoxycholic acid on cancer,dysplasia and mortality in ulcerative colitis patients with primary sclerosing cholangitis[J].Alimentary Pharmacology and Therapeutics,2005,22(9):783.
[4]黄晓燕,张涛,王囡囡,等.端粒缩短介导肠上皮细胞凋亡参与溃疡性结肠炎以及四君子汤干预研究[J].时珍国医国药,2014,25(9):2303.
[5]MANTOVANI A,ALLAVENA P,SICA A,et al.Cancer-related inflammation[J].Nature,2008,454(7203):436.
[6]COLOTTA F,ALLAVENA P,SICA A,et al.Cancer-related inflammation,the seventh hallmark of cancer:links to genetic instability[J].Carcinogenesis,2009,30(7):1073.
[7]李仪奎.中药药理实验方法学[M].2版.上海:上海科学技术出版社,2006:24.
[8]施胜英,林海桢,周溦,等.加味四君子汤含药血清对肝癌Hep-G2细胞的影响[J].中国实验方剂学杂志,2016,22(18):88.
[9]李景南.溃疡性结肠炎癌变机制的研究进展:从炎症到肿瘤[J].中华消化杂志,2011,31(3):183.
[10]SHORT B.Mitochondrial caspase keeps autophagy in flux[J].The Journal of Cell Biology,2014,205(4):431.
[11]于卫华,周庆彪,刘颖,等.活性氧调控炎症诱发肿瘤机制的研究进展[J].癌变·畸变·突变,2016,28(2):158.
[12]SHOJI Y,YOSHINAGA K,INOUE A,et al.Quantification of telomerase activity in sporadic colorectal carcinoma[J].Cancer,2000,88(6):1304.
[13]谢裕安,葛莲英,刘剑仑,等.端粒酶在大肠癌组织中表达的临床意义[J].现代肿瘤医学,2006,14(2):186.
[14]SUN J W,FAN L J,LI M N,et al.Decreasing Pin1suppresses telomerase activity by NF-κB in HCT116cells colorectal carcinoma[J].The Chinese-German Journal of Clinical Oncology,2013,12(4):181.
[15]KLEIDEITER E,FRIEDRICH U,M?HRING A,et al.Telomerase activity in chronic inflammatory bowel disease[J].Digestive Diseases and Sciences,2003,48(12):2328.
[16]葛莲英,刘爱群,罗小玲,等.RNA干扰沉默h TERT基因对大肠癌SW480细胞生物学特性的影响[J].中华实验外科杂志,2012,29(2):195.1979