Study On The Mechanism Of Saikosaponin D In Invasive Pituitary Adenoma Based On Long-chain Non-coding RNA KCNQ1OT1

Objective: To observe and compare the effects of different expressions of Long-chain non-coding RNA KCNQ1OT1(lnc RNA KCNQ1OT1)on cell proliferation,invasion and migration in rat pituitary adenoma GH3 cell lines.Meanwhile,the intervention mechanism of Saikosaponin D(SSD)on GH3 cell line of rat pituitary adenoma and the effect of SSD on the expression of lnc RNA KCNQ1OT1 in GH3 cell line of rat pituitary adenoma were studied.Objective: To investigate the regulation of SSD through its target gene lnc RNA KCNQ1OT1 on the invasion progression of GH3 cells,and to provide experimental basis and theoretical experience for the early diagnosis and subsequent treatment of invasive PAs in clinical practice.Methods: The constructed pc DNA3.1-KCNQ1OT1 plasmid and the negative control pc DNA3.1-KCNQ1OT1 NC were transfected into GH3 cells of rat pituitary adenoma to obtain KCNQ1OT1 group and KCNQ1OT1 NC group,respectively.SSDS were dissolved in dimethyl sulfoxide and added to GH3 cell medium and KCNQ1OT1 group for further culture,which were named SSD group and KCNQ1OT1+SSD group,respectively.Then,total RNA was extracted from GH3 cells of rat pituitary adenoma.Reverse transcription reaction and real-time quantitative PCR were performed to detect different expressions of target genes in KCNQ1OT1+SSD group,KCNQ1OT1 group and KCNQ1OT1 NC group.Finally,cell function tests,including CCK-8 assay,clonogenesis assay and transwell assay,were conducted to compare the number of proliferation,clonogenesis and invasion cells of GH3 cells after overexpression of KCNQ1OT1 and GH3 cells under SSD.And the changes of the number of GH3 cell proliferation,clone formation and invasion under the effect of SSD on KCNQ1OT1.Results: Real-time quantitative PCR showed that KCNQ1OT1 expression was significantly up-regulated in lnc RNA KCNQ1OT1 group,and KCNQ1OT1 gene expression was significantly down-regulated in GH3 of rat pituitary cells in lnc RNA KCNQ1OT1+SSD group(P < 0.001).After acting on GH3 cells of pituitary adenoma,SSD can significantly inhibit cell proliferation,clonogenesis and cell invasiveness.CCK-8 assay showed that the number of living cells in lnc RNA KCNQ1OT1 group increased significantly at 48h(P < 0.05),72h(P < 0.01)and96h(P < 0.05).The number of living cells in SSD group was significantly decreased at 48h(P < 0.001),72h(P < 0.01)and 96h(P < 0.001),and the number of living cells in lnc RNA KCNQ1OT1+SSD group was significantly decreased at48 h,72h and 96h(P < 0.05).Cell clonogenesis test and cell invasion test showed that the number of clonogenesis and invasion in lnc RNA KCNQ1OT1 group was significantly increased(P < 0.001),the number of clonogenesis and invasion in SSD group was significantly decreased,and the number of invasive cells in lnc RNA KCNQ1OT1+SDD group was significantly decreased(P < 0.01).Conclusions: lnc RNA KCNQ1OT1 is highly expressed in GH3 of rat pituitary tumor cells and promotes its proliferation,clonogenesis and invasion.SSD can improve the proliferation,clonogenesis and invasion of GH3 of rat pituitary tumor cells by inhibiting the overexpression of lnc RNA KCNQ1OT1.