重楼皂苷Ⅶ调控BBOX1-AS1对肾癌细胞的影响

PolyphyllinⅦregulate the effect of BBOX1-AS1 on renal cancer cells

目的探讨重楼皂苷Ⅶ(PP7)通过调控长链非编码RNA(lncRNA)BBOX1反义RNA 1(BBOX1-AS1)对肾癌细胞增殖、迁移和侵袭的影响。方法将肾癌786-0细胞分为对照组、0.1μmol/L、0.3μmol/L、1μmol/L重楼皂苷Ⅶ(PP7-L组、PP7-M组、PP7-H组)、sh-LV(转染sh-LV)组、sh-BBOX1-AS1-LV(转染sh-BBOX1-AS1-LV)组、BBOX1-AS1-LV(转染pcDNA 3.1 BBOX1-AS1)组、PP7-H+BBOX1-AS1-LV(1μmol/L重楼皂苷Ⅶ+转染pcDNA 3.1 BBOX1-AS1)组。运用集落形成实验检测细胞集落形成能力,MTT法检测细胞活性,划痕试验检测细胞迁移,Transwell实验检测细胞侵袭,蛋白质印迹法检测上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)的表达,实时逆转录聚合酶链反应检测BBOX1-AS1表达水平。结果与对照组相比,PP7-L、PP7-M和PP7-H组集落形成数、细胞活性、划痕愈合率、侵袭细胞数、N-cadherin蛋白表达和BBOX1-AS1表达水平逐渐降低,E-cadherin蛋白表达逐渐增加,差异均有统计学意义(P<0.05),并呈浓度依赖性。干扰BBOX1-AS1降低786-0细胞的划痕愈合率、侵袭细胞数、细胞活性、集落形成数、BBOX1-AS1表达水平、N-cadherin蛋白表达,提高E-cadherin表达水平,差异均有统计学意义(P<0.05)。过表达BBOX1-AS1逆转重楼皂苷Ⅶ对786-0增殖迁移侵袭的影响。结论重楼皂苷Ⅶ通过下调BBOX1-AS1,抑制肾癌细胞的增殖、迁移和侵袭。

Objective To investigate the effects of polyphyllinⅦ(PP7)on the proliferation,migration and invasion of renal cancer cells by regulating long non-coding RNA(lncRNA)BBOX1 antisense RNA 1(BBOX1-AS1).Methods Divided renal cell carcinoma 786-0 cells into control group,0.1μmol/L,0.3μmol/L,1μmol/L polyphyllinⅦ(PP7-L group,PP7-M group,PP7-H group),sh-LV(transfection sh-LV)group,sh-BBOX1-AS1-LV(transfection sh-BBOX1-AS1-LV)group,BBOX1-AS1-LV(transfection pcDNA 3.1 BBOX1-AS1)group,PP7-H+BBOX1-AS1-LV(1μmol/L saponinsⅦ+transfected with pcDNA 3.1 BBOX1-AS1)group.Colony formation test was used to detect cell colony forming ability,MTT method determined cell viability,scratch test was used to detect cell migration,and Transwell test detected cell invasion,Western blotting to detect the expression of epithelial cadherin(E-cadherin)and neural cadherin(N-cadherin).Real-time reverse transcription polymerase chain reaction detected the expression level of BBOX1-AS1.Results Compared with the control group,the number of colonies formed,cell viability,scratch healing rate,number of invasive cells,N-cadherin protein expression and BBOX1-AS1 expression levels in PP7-L group,PP7-M group and PP7-H group gradually decreased,and E-cadherin protein expression gradually increased,and the differences were statistically significant(P<0.05),which was concentration-dependent.Interfering with BBOX1-AS1 reduced the scratch healing rate,number of invasive cells,cell viability,number of colonies formed,BBOX1-AS1 expression level,N-cadherin protein expression,and increased E-cadherin expression level of 786-0 cells,the differences were statistically significant(P<0.05).Overexpression of BBOX1-AS1 reversed the effect of polyphyllinⅦon the proliferation,migration and invasion of 786-0.Conclusion PolyphyllinⅦdown-regulated BBOX1-AS1 to inhibit the proliferation,migration and invasion of renal cancer cells.