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β-榄香烯通过调控长链非编码RNA介导的hTERT表达抑制来抑制食管鳞状细胞癌的增殖。

β-Elemene inhibits the proliferation of esophageal squamous cell carcinoma by regulating long noncoding RNA-mediated inhibition of hTERT expression.

作者信息

Hu Zhaoyang, Wu Hongjin, Li Ying, Hou Qiang, Wang Yan, Li Shuang, Xia Bing, Wu Shixiu

机构信息

Hangzhou Cancer Institute, Hangzhou Cancer Hospital, Hangzhou City, China.

出版信息

Anticancer Drugs. 2015 Jun;26(5):531-9. doi: 10.1097/CAD.0000000000000216.

Abstract

The study aimed to clarify the relationship between β-elemene, a long noncoding RNA (lncRNA), and human telomerase reverse transcriptase (hTERT) in esophageal carcinoma ECA-109 cells. The proliferation of ECA-109 cells was measured using a CCK-8 kit and flow cytometry. PCR microarray and real-time RT-PCR were designed to determine lncRNA expression in ECA-109 cells before and after treatment with β-elemene. Western blot was used to detect the hTERT level after the differentially expressed lncRNAs in ECA-109 cells were interfered with small interfering RNA (siRNA). On treatment with β-elemene, the proliferation of ECA-109 cells was notably inhibited, and about 85% of the lncRNAs showed higher expression levels in ECA-109 cells than in those untreated cells, from which, CDKN2B-AS1 was screened out. A specific siRNA (si-CDKN2B-AS1) that targets the β-elemene-mediated lncRNA CDKN2B-AS1 was designed, synthesized, and applied to treat ECA-109 cells. Its interference efficiency reached as high as 89.6%. When ECA-109 cells were transfected with the siRNA, the hTERT level was increased by 84.7%. The CCK-8 assay showed that the proliferation of ECA-109 cells treated with β-elemene was significantly promoted after siRNA transfection (P<0.01). It was also shown by flow cytometry that, compared with the scramble-treated group (negative control), the proliferation index value of ECA-109 cells in the si-CDKN2B-AS1 treatment group was notably increased (25.7 vs. 51.7%) and the TERT protein level was increased by 67.25% after the cells were treated with si-CDKN2B-AS1. The chemotherapeutic drug β-elemene suppressed the proliferation of esophageal carcinoma ECA-109 cells by regulating the inhibition of hTERT expression by lncRNA CDKN2B-AS1.

摘要

该研究旨在阐明长链非编码RNA(lncRNA)β-榄香烯与食管癌ECA-109细胞中人端粒酶逆转录酶(hTERT)之间的关系。使用CCK-8试剂盒和流式细胞术检测ECA-109细胞的增殖情况。设计PCR微阵列和实时逆转录PCR以确定β-榄香烯处理前后ECA-109细胞中lncRNA的表达。在ECA-109细胞中差异表达的lncRNAs被小干扰RNA(siRNA)干扰后,使用蛋白质免疫印迹法检测hTERT水平。用β-榄香烯处理后,ECA-109细胞的增殖受到显著抑制,约85%的lncRNAs在ECA-109细胞中的表达水平高于未处理细胞,从中筛选出CDKN2B-AS1。设计、合成了一种靶向β-榄香烯介导的lncRNA CDKN2B-AS1的特异性siRNA(si-CDKN2B-AS1),并将其应用于处理ECA-109细胞。其干扰效率高达89.6%。当用siRNA转染ECA-109细胞时,hTERT水平提高了84.7%。CCK-8检测表明,siRNA转染后,用β-榄香烯处理的ECA-109细胞的增殖得到显著促进(P<0.01)。流式细胞术也显示,与乱序处理组(阴性对照)相比,si-CDKN2B-AS1处理组中ECA-109细胞的增殖指数值显著增加(25.7%对51.7%),并且在用si-CDKN2B-AS1处理细胞后,TERT蛋白水平提高了67.25%。化疗药物β-榄香烯通过调节lncRNA CDKN2B-AS1对hTERT表达的抑制作用来抑制食管癌ECA-109细胞的增殖。

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