Zhang Xian-Juan, Qi Guang-Tao, Zhang Xiao-Min, Wang Li, Li Fang-Fang
Binzhou Medical University Hospital, Binzhou, Shandong, China.
J Obstet Gynaecol Res. 2021 Feb;47(2):533-543. doi: 10.1111/jog.14548. Epub 2020 Nov 9.
This study aimed to investigate the function of long noncoding RNA RHPN1 antisense RNA 1 (lncRNA RHPN1-AS1) in the progression of endometrial cancer (EC) and its underlying molecular mechanisms.
The RHPN1-AS1 expression was measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) in EC tissues and cells. The cell clones, proliferation, cell cycle, apoptosis, migration and invasion in Ishikawa and HEC-1A cells were respectively measured by colony formation assay, cell counting kit-8 assay (CCK-8) assay, flow cytometry, wound healing assay and transwell assay. In addition, the protein expressions in Ishikawa and HEC-1A cells were measured using western blot and Immunofluorescence assay.
Our data showed the RHPN1-AS1 expression was significantly upregulated in both EC tissues and cells. The expression of RHPN1-AS1 was significantly correlated with FIGO stage, histological grade, and lymph node metastasis. Additionally, silencing RHPN1-AS1 could inhibit proliferation, cell cycle progression, migration and invasion, and also promote apoptosis in Ishikawa and HEC-1A cells. Moreover, silencing RHPN1-AS1 could markedly elevate the expressions of caspase-3 and Bax, but reduce the Bcl-2 expression in Ishikawa and HEC-1A cells. We also found that silencing RHPN1-AS1 could significantly inhibit the phosphorylation of MEK and ERK in Ishikawa and HEC-1A cells. After U0126 pretreatment, the inhibition effect of silencing RHPN1-AS1 on the phosphorylation of MEK and ERK was strengthened.
Our study demonstrated that RHPN1-AS1 could facilitate cell proliferation, migration and invasion, as well as inhibit apoptosis via activating ERK/MAPK pathway in EC.
本研究旨在探讨长链非编码RNA RHPN1反义RNA 1（lncRNA RHPN1-AS1）在子宫内膜癌（EC）进展中的作用及其潜在分子机制。
采用定量逆转录聚合酶链反应（qRT-PCR）检测EC组织和细胞中RHPN1-AS1的表达。通过集落形成试验、细胞计数试剂盒-8试验（CCK-8）、流式细胞术、伤口愈合试验和Transwell试验分别检测Ishikawa和HEC-1A细胞的细胞克隆、增殖、细胞周期、凋亡、迁移和侵袭。此外，采用蛋白质印迹法和免疫荧光试验检测Ishikawa和HEC-1A细胞中的蛋白质表达。
我们的数据显示，RHPN1-AS1在EC组织和细胞中均显著上调。RHPN1-AS1的表达与国际妇产科联盟（FIGO）分期、组织学分级和淋巴结转移显著相关。此外，沉默RHPN1-AS1可抑制Ishikawa和HEC-1A细胞的增殖、细胞周期进程、迁移和侵袭，并促进其凋亡。此外，沉默RHPN1-AS1可显著提高Ishikawa和HEC-1A细胞中caspase-3和Bax的表达，但降低Bcl-2的表达。我们还发现，沉默RHPN1-AS1可显著抑制Ishikawa和HEC-1A细胞中MEK和ERK的磷酸化。U0126预处理后，沉默RHPN1-AS1对MEK和ERK磷酸化的抑制作用增强。
我们的研究表明，RHPN1-AS1可通过激活EC中的ERK/MAPK途径促进细胞增殖、迁移和侵袭，并抑制细胞凋亡。
