Effects of SERPINE1 on Proliferation and Apoptosis of Colon Adenocarcinoma Cells RKO and LoVo

CHEN Dehe; LI Dengyu; GUO Gang; WANG Jie; ZHOU Qian; LI Hongtao

doi:10.13730/j.issn.2097-2148.2024.04.002

Abstract：Objective To explore the expression and significance of serine proteinase inhibitor family E member 1(SERPINE1)in colon adenocarcinoma(COAD)and its effects on proliferation and apoptosis of COAD cells,and to ex-plore its biological function in the progression of COAD.Methods The relationship between SERPINE1 expression and clinical stage and prognosis of COAD was analyzed by Gene Expression Profilling Interactive Analysis 2.0(GEPIA 2.0)database.Cancer tissues and corresponding normal colonic tissues were collected from 5 patients with COAD,the expres-sion of SERPINE1 in COAD was verified by Western blot;then SERPINE1 overexpression and interference lentiviral vec-tors were constructed for cell transfection,real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to screen the experi-mental cells and verify the overexpression and interference expression efficiency;cell proliferation and apoptosis were detected by cell counting kit 8(CCK-8)and Hoechst nu-clear staining;finally,the effects of the inhibitor on the marker of proliferation Ki-67(MKI67),Bcl-2 associated X protein(Bax)and B cell lymphoma 2(Bcl-2)gene ratio were detected by Western blot.Results The GEPIA2 database was used to search and analyze the relationship between the expression of SERPINE1 in COAD and clinical stage as well as prognosis,it was found that the expression level of SERPINE1 in COAD was higher than that in normal colonic tis-sues,the expression level of SERPINE1 was positively correlated with clinical stage,but negatively correlated with prog-nosis and overall survival(all P＜0.05).The results of CCK-8 assay showed that the optical density(OD)value of COAD cells in oe-RKO group and oe-LoVo group at 48h was higher than that of oe-RKO-NC group and oe-LoVo-NC group,the OD value of COAD cells in sh-RKO-822 group and sh-LoVo-822 group at 48h was lower than that of sh-RKO-NC and sh-LoVo-NC groups(all P＜0.05).Hoechst staining showed that the number of apoptotic COAD cells in oe-RKO group and oe-LoVo group was lower than that in oe-RKO-NC group and oe-LoVo-NC group,the number of apoptotic COAD cells in sh-RKO-822 group and sh-LoVo-822 group was significantly higher than that in sh-RKO-NC group and sh-LoVo-NC group(all P＜0.05).Western blot results showed that the expression of COAD cells in MKI67 protein in oe-RKO group and oe-LoVo group was significantly higher than that in oe-RKO-NC group and oe-LoVo-NC group,and the Bax/Bcl-2 was significantly lower than that in oe-RKO-NC group and oe-LoVo-NC group;the expression of COAD cells in MKI67 protein in sh-RKO-822 group and sh-LoVo-822 group was significantly lower than that in sh-RKO-NC group and sh-LoVo-NC group,and the Bax/Bcl-2 was significantly higher than that in sh-RKO-NC group and sh-LoVo-NC group(all P＜0.05).Conclusion SERPINE1 is highly expressed in COAD and is positively correlated with clinical stage,but nega-tively correlated with prognosis;SERPINE1 may promote the proliferation and inhibit the apoptosis of COAD RKO and LoVo cells by affecting the expression of COAD cells in MKI67 and Bax/Bcl-2.