Abstract:

Objective To investigate the expression of long non-coding RNA（lncRNA）heart and neural crest derivatives expressed 2-antisense RNA 1（HAND2-AS1）in serum of cervical cancer patients and the mechanism of its effect on the proliferation，invasion and migration of cervical cancer Caski cells. Methods The Gene Expression Profiling Interactive Analysis（GEPIA） database was used to analyze the expression of lncRNA HAND2-AS1 in cervical cancer tissues and normal cervical tissues. From January 2019 to December 2021，48 patients with cervical cancer（cervical cancer group），48 patients with cervical intraepithelial neoplasia（CIN）（CIN group） and 48 healthy subjects（healthy control group） were enrolled from Jiaozuo Maternal and Child Health Care Hospital. The samples of postoperative cancer tissue and adjacent tissue（>2 cm from the margin of cancer tissue） were collected from 14 patients with cervical cancer. Serum samples and clinical data of all the subjects were collected to determine the relative expression level of lncRNA HAND2-AS1. Cervical cancer cell line Caski was classified into overexpression group（transfected with pcDNA3.1-HAND2-AS1），negative control group（transfected with pcDNA3.1-NC），interference group（transfected with si-HAND2-AS1） and interference control group（transfected with si-NC） according to different transfected plasmids. The proliferation，invasion and migration were determined by CCK-8 assay，Transwell assay and scratch test. The expressions of phosphatidylinositol 3-kinase（PI3K），phosphorylated phosphatidylinositol 3-kinase（p-PI3K），protein kinase B（Akt） and phosphorylated protein kinase B（p-Akt） were determined by western blot. Results The lncRNA HAND2-AS1 expression in cervical cancer tissues was lower than that in normal cervical epithelial tissues（P<0.05）. The relative expression level of lncRNA HAND2-AS1 in cancer tissues of 14 patients with cervical cancer was lower than that in adjacent tissues（P<0.01）. The relative expression level of serum lncRNA HAND2-AS1 in cervical cancer group was lower than those in CIN group and healthy control group（P<0.01），but there was no statistical significance between CIN group and healthy control group（P>0.05）. The relative expression level of serum lncRNA HAND2-AS1 had statistical significance among cervical cancer patients with different tumor sizes，the International Federation of Gynecology and Obstetrics（FIGO） stages and lymphatic metastasis（P<0.05）. The relative expression level of lncRNA HAND2-AS1 in overexpression group was higher than that in negative control group（P<0.01），while the cell proliferation activity，the number of invasive cells，scratch healing rate and the relative expression levels of p-PI3K and p-Akt protein in overexpression group were lower than those in negative control group（P<0.05）. The relative expression level of lncRNA HAND2-AS1 in interference group was lower than that in interference control group（P<0.01），and the cell proliferation activity，the number of invasive cells，scratch healing rate and the relative expression levels of p-PI3K and p-Akt protein in interference group were higher than those in interference control group（P<0.05）. There was no statistical significance in the relative expression levels of PI3K and Akt between overexpression group and negative control group，or between interference group and interference control group（P>0.05）. Conclusions Serum lncRNA HAND2-AS1 expression is low in patients with cervical cancer，and it is related to disease severity. The lncRNA HAND2-AS1 may affect the proliferation，invasion and migration of cervical cancer cells by regulating the PI3K/Akt signaling pathway.

Key words: Heart and neural crest derivatives expressed 2-antisense RNA 1, Long non-coding RNA, Cervical cancer, Invasion, Migration