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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