Department of Pathophysiology, Medical College of Qinghai University, Qinghai, P.R. China.
Key Laboratory of Hydatidosis Research, Qinghai University Affiliated Hospital, Qinghai, P.R. China.
Syst Biol Reprod Med. 2022 Oct-Dec;68(5-6):370-383. doi: 10.1080/19396368.2022.2101961. Epub 2022 Aug 25.
This study investigated the expression and clinical significance of long intergenic noncoding RNA 00665 (LINC00665) in ovarian cancer (OC), as well as its effect on the malignant biological behavior of OC cells. The expression of LINC00665, miR-148b-3p, and Krüppel-like factor 5 (KLF5) in OC tissues and cells were determined by RT-qPCR. Western blot was used to detect the protein expression of KLF5. The expression patterns of LINC00665 in nuclear and cytoplasm fractions were undertaken using RT-qPCR. In addition, CCK-8 assay, clone formation assay, transwell, scratch test, and flow cytometry were respectively used to detect the cell activity, proliferation, invasiveness, healing of cells, and apoptosis rate of OC cells. Furthermore, the interactions between LINC00665 and miR-148b-3p and between miR-148b-3p and KLF5 were verified by the luciferase reporter assay, and the correlations among these three genes were analyzed. LINC00665 expression was upregulated both in OC cell lines and tissues. Si-LINC00665 inhibited cell proliferation, invasion, and migration and induced apoptosis to a certain extent. The subcellular fraction assay revealed LINC00665 to be located mainly in the cytoplasm. miR-148b-3p was a target of LINC00665, and KLF5 was directly targeted by miR-148b-3p. Si-LINC00665 inhibited KLF5 expression, miR-148b-3p inhibitor promoted KLF5 expression, and si-KLF5 inhibited LINC00665 expression. Interestingly, the expression of LINC00665 was reversely associated with miR-148b-3p expression but positively correlated with KLF5. Furthermore, miR-148b-3p expression was negatively correlated with KLF5. In addition, si-KLF5 inhibited the malignant biological behavior of OC cells, whereas miR-148b-3p inhibitor had the opposite effect. Most importantly, the si-LINC00665 could reverse the promotion effect of the miR-148b-3p inhibitor on the malignant biological behavior of OC cells. LINC00665 can be used as an effective prognostic indicator of OC, which has the potential to be a new therapeutic target.
本研究探讨了长链非编码 RNA 00665(LINC00665)在卵巢癌(OC)中的表达及临床意义,以及其对 OC 细胞恶性生物学行为的影响。采用 RT-qPCR 检测 OC 组织和细胞中 LINC00665、miR-148b-3p 和 Krüppel 样因子 5(KLF5)的表达。采用 Western blot 检测 KLF5 蛋白表达。采用 RT-qPCR 检测核和胞质 LINC00665 的表达模式。分别采用 CCK-8 检测、克隆形成实验、Transwell、划痕实验和流式细胞术检测 OC 细胞的细胞活性、增殖、侵袭、细胞愈合和细胞凋亡率。此外,通过荧光素酶报告实验验证 LINC00665 与 miR-148b-3p 以及 miR-148b-3p 与 KLF5 之间的相互作用,并分析这三个基因之间的相关性。LINC00665 在 OC 细胞系和组织中均呈高表达。Si-LINC00665 可在一定程度上抑制细胞增殖、侵袭和迁移,诱导细胞凋亡。亚细胞分馏试验表明 LINC00665 主要位于细胞质中。miR-148b-3p 是 LINC00665 的靶标,而 KLF5 是 miR-148b-3p 的直接靶标。Si-LINC00665 抑制 KLF5 表达,miR-148b-3p 抑制剂促进 KLF5 表达,而 si-KLF5 抑制 LINC00665 表达。有趣的是,LINC00665 的表达与 miR-148b-3p 的表达呈负相关,而与 KLF5 的表达呈正相关。此外,miR-148b-3p 的表达与 KLF5 的表达呈负相关。此外,si-KLF5 抑制 OC 细胞的恶性生物学行为,而 miR-148b-3p 抑制剂则产生相反的效果。最重要的是,si-LINC00665 可以逆转 miR-148b-3p 抑制剂对 OC 细胞恶性生物学行为的促进作用。LINC00665 可作为 OC 的有效预后指标,具有成为新的治疗靶点的潜力。