LINC00665 affects the malignant biological behavior of ovarian cancer via the miR-148b-3p/KLF5.

IF 2.1 4区 医学 Q3 ANDROLOGY
Shenglan Wang, Chuanchuan Liu, Yongchuan Li, Jinwan Qiao, Xinling Chen, Jin Bao, Ran Li, Yanxia Xing
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Abstract

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.

LINC00665通过miR-148b-3p/KLF5影响卵巢癌的恶性生物学行为。
本研究探讨长基因间非编码RNA 00665 (LINC00665)在卵巢癌(OC)中的表达及临床意义,以及其对卵巢癌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、kr pel样因子5 (KLF5)的表达。Western blot检测KLF5蛋白表达。采用RT-qPCR检测LINC00665在细胞核和细胞质部分的表达谱。分别采用CCK-8法、克隆形成法、transwell法、划痕法、流式细胞术检测OC细胞的细胞活性、增殖、侵袭性、细胞愈合和凋亡率。此外,通过荧光素酶报告基因实验验证了LINC00665与miR-148b-3p、miR-148b-3p与KLF5之间的相互作用,并分析了这三个基因之间的相关性。在OC细胞系和组织中,LINC00665的表达均上调。Si-LINC00665在一定程度上抑制细胞增殖、侵袭和迁移,并诱导细胞凋亡。亚细胞组分分析显示LINC00665主要位于细胞质中。miR-148b-3p是LINC00665的靶点,而KLF5被miR-148b-3p直接靶向。Si-LINC00665抑制KLF5表达,miR-148b-3p抑制剂促进KLF5表达,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呈正相关。此外,miR-148b-3p表达与KLF5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抑制剂具有相反的作用。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,有潜力成为新的治疗靶点。
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来源期刊
CiteScore
4.30
自引率
4.20%
发文量
27
审稿时长
>12 weeks
期刊介绍: Systems Biology in Reproductive Medicine, SBiRM, publishes Research Articles, Communications, Applications Notes that include protocols a Clinical Corner that includes case reports, Review Articles and Hypotheses and Letters to the Editor on human and animal reproduction. The journal will highlight the use of systems approaches including genomic, cellular, proteomic, metabolomic, bioinformatic, molecular, and biochemical, to address fundamental questions in reproductive biology, reproductive medicine, and translational research. The journal publishes research involving human and animal gametes, stem cells, developmental biology and toxicology, and clinical care in reproductive medicine. Specific areas of interest to the journal include: male factor infertility and germ cell biology, reproductive technologies (gamete micro-manipulation and cryopreservation, in vitro fertilization/embryo transfer (IVF/ET) and contraception. Research that is directed towards developing new or enhanced technologies for clinical medicine or scientific research in reproduction is of significant interest to the journal.
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