摘要
目的 探讨虎杖苷调节环磷酸腺苷(cAMP)/环磷酸腺苷结合蛋白1(EPAC1)/Ras相关蛋白1(RAP1)信号通路对子宫内膜癌细胞恶性生物学行为的影响。方法 研究时间为2022年8月-2023年8月。以子宫内膜癌Ishikawa细胞为研究对象,分为空白组、虎杖苷-L组(25μmol/L虎杖苷)、虎杖苷-M组(50μmol/L虎杖苷)、虎杖苷-H组(100μmol/L虎杖苷)、Forskolin组(cAMP激活剂,100μmol/L Forskolin)以及虎杖苷-H+Forskolin组(100μmol/L虎杖苷+100μmol/L Forskolin)。分别以克隆形成实验、Transwell实验、划痕实验、TUNEL法、CCK-8法、Western Blot检测细胞克隆数、侵袭、迁移、凋亡、增殖及EPAC1、RAP1、凋亡蛋白Cleaved Caspase-3、基质金属蛋白酶-9(MMP-9)蛋白表达。采用ELISA法检测细胞培养基中cAMP水平。结果 与空白组相比,Forskolin组细胞克隆数[(195.66±19.62)个]、侵袭数[(281.52±28.22)个]、迁移率[(82.61±8.31)%]、增殖率[(125.64±12.59)%]、cAMP水平[(1 385.54±138.56)pmol/ml]、EPAC1(1.56±0.16)、RAP1(2.11±0.22)、MMP-9(1.88±0.19)表达显著增加,凋亡率[(5.47±0.56)%]、Cleaved Caspase-3(0.26±0.03)表达均降低(均P<0.05),虎杖苷-L组、虎杖苷-M组、虎杖苷-H组细胞克隆数[(105.34±10.57)个、(65.34±6.58)个、(42.27±4.25)个]、侵袭数[(185.72±18.66)个、(145.47±14.58)个、(112.59±11.29)个]、迁移率[(40.21±4.06)%、(31.12±3.12)%、(21.34±2.15)%]、增殖率[(68.37±6.86)%、(48.67±4.88)%、(35.24±3.55)%]、cAMP水平、EPAC1、RAP1、MMP-9表达显著降低,凋亡率[(19.85±1.99)%、(27.34±2.77)%、(36.44±3.66)%]、Cleaved Caspase-3表达均增加(均P<0.05)。与Forskolin组相比,虎杖苷-H+Forskolin组细胞克隆数、侵袭数、迁移率、增殖率、cAMP水平、EPAC1、RAP1、MMP-9表达均显著降低,凋亡率、Cleaved Caspase-3表达均显著增加(均P<0.05)。与虎杖苷-H组相比,虎杖苷-H+Forskolin组细胞克隆数、侵袭数、迁移率、cAMP水平、EPAC1、RAP1、MMP-9表达均显著增加,凋亡率、增殖率、Cleaved Caspase-3表达均显著降低(均P<0.05)。结论 虎杖苷可抑制子宫内膜癌细胞恶性生物学行为发展,可能与抑制cAMP/EPAC1/RAP1信号通路有关。
Objective To explore the effect of polydatin on malignant biological behavior of endometrial cancer cells by regulating cy-clic adenosine monophosphate(cAMP)/cyclic adenosine phosphate binding protein(EPAC1)/Ras-associated protein 1(RAP1)signa-ling pathway.Methods The research time was from August 2022 to August 2023.Endometrial cancer Ishikawa cells were used as the re-search object,and they were divided into blank group,polydatin-L group(25 μmol/L polydatin),polydatin-M group(50 μmol/L polyda-tin),polydatin-H group(100 μmol/L polydatin),Forskolin group(cAMP activator,100 μmol/L Forskolin),and polydatin-H+Forskolin group(100 μmol/L polydatin and 100 μmol/L Forskolin).Clone formation experiment,Transwell experiment,scratch experiment,TUNEL method,CCK-8 method,and Western blot were used to detect cell clone number,invasion,migration,apoptosis,proliferation,and expressions of EPAC1,RAP1,apoptotic protein Cleaved Caspase-3,and matrix metalloproteinase-9(MMP-9).ELISA was used to de-tect cAMP level in cell culture media.Results Compared with blank group,cell clone count(195.66±19.62),number of invasion(281.52±28.22),migration rate[(82.61±8.31)%],proliferation rate[(125.64±12.59)%],the level of cAMP[(1 385.54±138.56)pmol/ml],the expression of EPAC1(1.56±0.16),RAP1(2.11±0.22),MMP-9(1.88±0.19)in Forskolin group increased significantly,the apoptosis rate[(5.47±0.56)%]and expression of Cleaved Caspase-3(0.26±0.03)decreased significantly(P<0.05),the cell clone counts[(105.34±10.57),(65.34±6.58),(42.27±4.25)],numbers of invasion[(185.72±18.66),(145.47±14.58),(112.59±11.29)],migration rates[(40.21±4.06)%,(31.12±3.12)%,(21.34±2.15)%],proliferation rates[(68.37±6.86)%,(48.67±4.88)%,(35.24±3.55)%],the levels of cAMP,the expressions of EPAC1,RAP1,and MMP-9 in poly-datin-L group,polydatin-M group,and polydatin-H group decreased significantly,the apoptosis rates[(19.85±1.99)%,(27.34±2.77)%,(36.44±3.66)%]and expressions of Cleaved Caspase-3 increased significantly(P<0.05).Compared with Forskolin group,the cell clone count,number of invasion,migration rate,proliferation rate,the level of cAMP,the expressions of EPAC1,RAP1,and MMP-9 in polydatin-H+Forskolin group decreased significantly,the apoptosis rate and expression of Cleaved Caspase-3 increased signifi-cantly(P<0.05).Compared with polydatin-H group,the cell clone count,number of invasion,migration rate,the level of cAMP,the ex-pressions of EPAC1,RAP1,and MMP-9 in polydatin-H+Forskolin group increased significantly,the apoptosis rate,proliferation rate,and expression of Cleaved Caspase-3 decreased significantly(P<0.05).Conclusion Polydatin can inhibit the development of malignant bio-logical behavior of endometrial cancer cells,which may be related to the inhibition of cAMP/EPAC1/RAP1 signaling pathway.
作者
刘佳
纪贤芬
严金金
LIU Jia;JI Xian-fen;YAN Jin-jin(Oncology Department,Jiangxi Maternal and Child Health Hospital,Nanchang,Jiangxi 330002,China)
出处
《中国妇幼保健》
CAS
2024年第15期2917-2921,共5页
Maternal and Child Health Care of China
基金
江西省中医药管理局科技计划课题(SZYYB20224630)。
