奥拉帕尼对卵巢癌SKOV3细胞PI3K/AKT/m TOR信号通路的影响
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摘要
目的分析奥拉帕尼抑制卵巢癌SKOV3细胞增殖的PI3K/AKT/mTOR信号通路机制。方法将生长状态良好的SK-OV3细胞随机分为5组:对照组、奥拉帕尼低剂量组(10 mg·L~(-1))、奥拉帕尼中剂量组(30 mg·L~(-1))、奥拉帕尼高剂量组(90mg·L~(-1))和PI3K蛋白抑制剂LY294002组(10 mg·L~(-1))。除对照组外,其余各组细胞给予相应剂量的化合物孵育,DAPI及免疫印迹法分析细胞凋亡蛋白的表达,q PCR、免疫印迹法和免疫细胞化学法测定PI3K/AKT/mTOR信号通路相关蛋白的表达。结果奥拉帕尼处理后卵巢癌SKOV3细胞增殖明显受到抑制(P<0.05),SKOV3细胞内的凋亡小体逐渐增多,促细胞凋亡蛋白caspase 3及caspase 9的表达逐步增强(P<0.05),PI3K、AKT和mTOR表达逐步降低(P<0.05),且随着奥拉帕尼水平的升高,作用越明显(P<0.05)。结论奥拉帕尼对卵巢癌SKOV3细胞具有剂量依赖性抑制作用,且可能是通过抑制PI3K/AKT/mTOR信号通路发挥作用。
OBJECTIVE To analyze the involvement of PI3K/AKT/mTOR signaling pathway on the inhibiting the proliferation of ovarian cancer SK-OV3 cell line by olaparib.METHODS The SKOV3 cells were randomly divided into five groups:control group,olaparib low dose group(10 mg·L~(-1)),olaparib medium dose group(30 mg·L~(-1)),olaparib high dose group(90 mg·L~(-1)),and PI3K protein inhibitor LY294002 group(10 mg·L~(-1)).In addition to the control group,the rest of the cells were incubated with the corresponding doses of the compounds.DAPI and Western blotting were used to analyze the expression of apoptotic protein.qPCR,Western blotting and immunocytochemistry were adopted to detect the expression of PI3K/AKT/mTOR signaling pathway related proteins.RESULTS After olaparib treatment,the proliferation of ovarian cancer SKOV3 cells was significantly inhibited(P<0.05).Apoptotic bodies in SKOV3 cells increased gradually,and the expression of caspase 3 and caspase 9 increased gradually(P<0.05),PI3K,AKT and mTOR expression gradually decreased(P<0.05)after olaparib medication.With the increase of olaparib level,the above effects were more obvious(P<0.05).CONCLUSION Olaparib has an inhibitory effect on ovarian cancer SKOV3 cells,and it may play a role by inhibiting the PI3K/AKT/mTOR signaling pathway.
OBJECTIVE To analyze the involvement of PI3K/AKT/mTOR signaling pathway on the inhibiting the proliferation of ovarian cancer SK-OV3 cell line by olaparib.METHODS The SKOV3 cells were randomly divided into five groups:control group,olaparib low dose group(10 mg·L~(-1)),olaparib medium dose group(30 mg·L~(-1)),olaparib high dose group(90 mg·L~(-1)),and PI3K protein inhibitor LY294002 group(10 mg·L~(-1)).In addition to the control group,the rest of the cells were incubated with the corresponding doses of the compounds.DAPI and Western blotting were used to analyze the expression of apoptotic protein.qPCR,Western blotting and immunocytochemistry were adopted to detect the expression of PI3K/AKT/mTOR signaling pathway related proteins.RESULTS After olaparib treatment,the proliferation of ovarian cancer SKOV3 cells was significantly inhibited(P<0.05).Apoptotic bodies in SKOV3 cells increased gradually,and the expression of caspase 3 and caspase 9 increased gradually(P<0.05),PI3K,AKT and mTOR expression gradually decreased(P<0.05)after olaparib medication.With the increase of olaparib level,the above effects were more obvious(P<0.05).CONCLUSION Olaparib has an inhibitory effect on ovarian cancer SKOV3 cells,and it may play a role by inhibiting the PI3K/AKT/mTOR signaling pathway.
引文
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[16]BAEK S H,KO J H,LEE J H,et al.Ginkgolic acid inhibits invasion and migration and tgf-beta-induced emt of lung cancer cells through pi3k/akt/mtor inactivation[J].J Cell Physiol,2017,232(2):346-354.
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[19]CAI Y,TAN X,LIU J,et al.Inhibition of pi3k/akt/mtor signaling pathway enhances the sensitivity of the skov3/ddp ovarian cancer cell line to cisplatin in vitro[J].Chin J Cancer Res(中国癌症研究),2014,26(5):564-572.
[20]BANG Y J,IM S A,LEE K W,et al.Randomized,doubleblind phase ii trial with prospective classification by atm protein level to evaluate the efficacy and tolerability of olaparib plus paclitaxel in patients with recurrent or metastatic gastric cancer[J].J Clin Oncol,2015,33(33):3858-3865.
[21]CHEDGY E C,BLACK P C.Moving beyond the androgen receptor in advanced prostate cancer commentary on:DNA-repair defects and olaparib in metastatic prostate cancer[J].Urology,2016,8910-11.
[22]VAN DER NOLL R,MARCHETTI S,STEEGHS N,et al.Longterm safety and anti-tumour activity of olaparib monotherapy after combination with carboplatin and paclitaxel in patients with advanced breast,ovarian or fallopian tube cancer[J].Br J Cancer,2015,113(3):396-402.
[2]HUANG L,JIN Y,FENG S,et al.Role of wnt/beta-catenin,wnt/c-jun n-terminal kinase and wnt/Ca2+pathways in cisplatininduced chemoresistance in ovarian cancer[J].Exp Ther Med,2016,12(6):3851-3858.
[3]HUANG R Y,ANTONY J,TAN T Z,et al.Targeting the axl signaling pathway in ovarian cancer[J].Mol Cell Oncol,2017,4(2):e1263716.
[4]LIU X,FENG M,ZHENG G,et al.Tcrp1 expression is associated with platinum sensitivity in human lung and ovarian cancer cells[J].Oncol Lett,2017,13(3):1398-1405.
[5]LU J,XU Y,WEI X,et al.Emodin inhibits the epithelial to mesenchymal transition of epithelial ovarian cancer cells via ilk/gsk-3beta/slug signaling pathway[J].Biomed Res Int,2016,20166253280.
[6]CHEAIB B,AUGUSTE A,LEARY A.The pi3k/akt/mtor pathway in ovarian cancer:therapeutic opportunities and challenges[J].Chin J Cancer(癌症),2015,34(1):4-16.
[7]YU P,YE L,WANG H,et al.Nsk-01105 inhibits proliferation and induces apoptosis of prostate cancer cells by blocking the raf/mek/erk and pi3 k/akt/mtor signal pathways[J].Tumour Biol,2015,36(3):2143-2153.
[8]PUJADE-LAURAINE E,COMBE P.Olaparib in ovarian cancer with brca mutation[J].Bull Cancer,2015,102(6 suppl 1):S82-84.
[9]TEWARI K S,ESKANDER R N,MONK B J.Development of olaparib for brca-deficient recurrent epithelial ovarian cancer[J].Clin Cancer Res,2015,21(17):3829-3835.
[10]JIANG C M H Z,YANG Q.Effect of apoptosis inducing of ginsenoside compound k on chronic myelocytic leukemia k562 cells and its mechanism[J].J Chin Oncol(肿瘤学杂志),2014,20(2):122-126.
[11]VAN DER STEEN S C,RAAVE R,LANGERAK S,et al.Targeting the extracellular matrix of ovarian cancer using functionalized,drug loaded lyophilisomes[J].Eur J Pharm Biopharm,2017,113229-239.
[12]ZHANG S,NG M K.Gene-microrna network module analysis for ovarian cancer[J].BMC Syst Biol,2016,10(suppl 4):117.
[13]BAI X,MA Y,ZHANG G.Butein suppresses cervical cancer growth through the pi3k/akt/mtor pathway[J].Oncol Rep,2015,33(6):3085-3092.
[14]CHENG H,SHCHERBA M,PENDURTI G,et al.Targeting the pi3k/akt/mtor pathway:potential for lung cancer treatment[J].Lung Cancer Manag,2014,3(1):67-75.
[15]PENG Y,LI L,HUANG M,et al.Angiogenin interacts with ribonuclease inhibitor regulating pi3k/akt/mtor signaling pathway in bladder cancer cells[J].Cell Signal,2014,26(12):2782-2792.
[16]BAEK S H,KO J H,LEE J H,et al.Ginkgolic acid inhibits invasion and migration and tgf-beta-induced emt of lung cancer cells through pi3k/akt/mtor inactivation[J].J Cell Physiol,2017,232(2):346-354.
[17]BLANCO E,SANGAI T,WU S,et al.Colocalized delivery of rapamycin and paclitaxel to tumors enhances synergistic targeting of the pi3k/akt/mtor pathway[J].Mol Ther,2014,22(7):1310-1319.
[18]BAI H,LI H,LI W,et al.The pi3k/akt/mtor pathway is a potential predictor of distinct invasive and migratory capacities in human ovarian cancer cell lines[J].Oncotarget,2015,6(28):25520-25532.
[19]CAI Y,TAN X,LIU J,et al.Inhibition of pi3k/akt/mtor signaling pathway enhances the sensitivity of the skov3/ddp ovarian cancer cell line to cisplatin in vitro[J].Chin J Cancer Res(中国癌症研究),2014,26(5):564-572.
[20]BANG Y J,IM S A,LEE K W,et al.Randomized,doubleblind phase ii trial with prospective classification by atm protein level to evaluate the efficacy and tolerability of olaparib plus paclitaxel in patients with recurrent or metastatic gastric cancer[J].J Clin Oncol,2015,33(33):3858-3865.
[21]CHEDGY E C,BLACK P C.Moving beyond the androgen receptor in advanced prostate cancer commentary on:DNA-repair defects and olaparib in metastatic prostate cancer[J].Urology,2016,8910-11.
[22]VAN DER NOLL R,MARCHETTI S,STEEGHS N,et al.Longterm safety and anti-tumour activity of olaparib monotherapy after combination with carboplatin and paclitaxel in patients with advanced breast,ovarian or fallopian tube cancer[J].Br J Cancer,2015,113(3):396-402.
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