长链非编码RNA-H19在妇科恶性肿瘤细胞的糖代谢中的作用

Functions of Long Noncoding RNA H19 in Glucose Metabolism of Gynecological Malignancy Cells

目的:检测长链非编码RNA-H19在常见的妇科恶性肿瘤中的表达,并探讨其在肿瘤细胞糖代谢过程中发挥的可能作用。方法:以ARK2子宫内膜癌细胞株和OVCAR-3上皮性卵巢癌细胞株作为实验对象;利用脂质体转染技术,将特异性靶向小干扰RNA序列转染进细胞;利用TRIzol法提取样品的总RNA,并通过逆转录和实时定量PCR技术检测样品中目的基因的表达;利用荧光标记的非放射性2-NBDG评估肿瘤细胞的葡萄糖摄取能力;采用方差齐性检验和t检验进行统计学分析。结果:H19在ARK2和OVCAR-3细胞中的表达均较高。在ARK2和OVCAR-3细胞中,si H19抑制H19表达的效率分别为34.02%和30.30%。是否添加Insulin对于转染si Con的肿瘤细胞的糖摄取能力无明显影响,但是si H19转染后的ARK2和OVCAR-3细胞,在经Insulin处理后糖摄取能力均明显增强。结论:ARK2和OVCAR-3这两种细胞是研究H19在妇科恶性肿瘤中机制的较为理想的实验对象,ARK2和OVCAR-3细胞本身存在胰岛素抵抗,外源性抑制H19表达后,细胞的胰岛素抵抗有所改善,H19或成为我们改善妇科恶性肿瘤患者机体葡萄糖代谢的标记物。

Objective： To investigate the expression of long noncoding RNA H19 in some kinds of common gynecological cancers and its correlation with glucose metabolism in these malignancies, and to explore the possible mechanism involved in the pathogenesis of gynecological malignancy. Methods： ARK2 endometrial cancer cell line and OVCAR-3 epithelial ovarian cancer cell line was used as objects. Lipofection was used to transfect some specific si RNAs into cancer cells. Total RNA extracted by TRIzol was used for target genes analysis through reverse transcription and then real time PCR. Glucose uptake assays were carried out using 2-NBDG, a fluorescently-labeled deoxyglucose analog, as a tracer for direct monitoring of glucose transport in live cells. The data were analyzed by homogeneity test of variance and t test. Results： Lnc RNA-H19 was highly expressed in both ARK2 and OVCAR-3 cells. H19 expressions could be inhibited successfully by RNA interference induced by si H19 in both two cell lines（interference rate as 34.02 % and 30.30 %respectively）. Importantly, there was no insulin-dependent stimulation of glucose uptake in both ARK2 and OVCAR-3 cells transfected with si Con. In contrast, insulin respectively stimulated glucose uptake by 1.42- and 1.60-fold in ARK2 and OVCAR-3 cells transfected with si H19. Conclusions： ARK2 and OVCAR-3 cell lines were ideal experimental objects to study the mechanism of H19 in gynecological cancers. Insulin resistance existed in both ARK2 and OVCAR-3 cells with highly expression of H19. This resistance could be greatly improved when H19 expression was suppressed. As the result, H19 might be an effective biomarker and can be used to repair glucose metabolism in gynecological malignancies in the future.