Raptor的表达与西罗莫司对膀胱癌BIU-87细胞周期及迁移的影响
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摘要
目的探讨Raptor在膀胱癌BIU-87细胞中的表达及西罗莫司对BIU-87细胞迁移和细胞周期的影响。方法将BIU-87细胞贴壁培养于10%胎牛血清、100 U/mL青霉素、100μg/mL链霉素的RPMI-1640培养液中,37℃、5%CO_2饱和湿度培养箱中培养。取对数生长期细胞,处理组加入浓度分别为(50-200)nmol/L西罗莫司;对照组不加药物,每天更换1640培养液。实时PCR法检测Raptor在BIU-87细胞中的表达,Transwell法检测BIU-87细胞的体外迁移能力,流式细胞仪检测不同浓度西罗莫司对BIU-87细胞周期的影响。结果西罗莫司处理组的Raptor的表达量明显下降,BIU-87体外迁移能力明显下降,呈剂量依赖性。西罗莫司将BIU-87细胞周期阻滞在G_0/G_1期,从而抑制了细胞的增殖。结论西罗莫司抑制BIU-87细胞增殖和迁移的能力与抑制Raptor的表达相关。
Objective To investigate the expression of Raptor in bladder cancer BIU-87 cells and the effect of sirolimus on the migration and cell cycle of BIU-87 cells. Methods BIU-87 cells were cultured in RPMI-1640 culture medium with 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin in 37℃ and 5%CO_2 saturated humidity incubator. Logarithmic growth phase cells were added to the treatment group with the concentration of(50-200)nmol/L sirolimus. Real-time PCR was used to measure the expression of Raptor in BIU-87 cells. Transwell assay was used to evaluate the ability of BIU-87 cells to migrate in vitro. Flow cytometry was used to detect the effect of sirolimus on the cell cycle of BIU-87 cells. Results The expression level of Raptor was significantly decreased, the migration ability of BIU-87 cells was significantly decreased in a dose-dependent manner, in sirolimus treated group than in the control. Sirolimus blocked BIU-87 cell cycle in G_0/G_1 phase, thereby inhibiting the cell proliferation. Conclusion The inhibition of Sirolimus on the proliferation and migration of BIU-87 cells might be related to inhibiting the expression of Raptor.
Objective To investigate the expression of Raptor in bladder cancer BIU-87 cells and the effect of sirolimus on the migration and cell cycle of BIU-87 cells. Methods BIU-87 cells were cultured in RPMI-1640 culture medium with 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin in 37℃ and 5%CO_2 saturated humidity incubator. Logarithmic growth phase cells were added to the treatment group with the concentration of(50-200)nmol/L sirolimus. Real-time PCR was used to measure the expression of Raptor in BIU-87 cells. Transwell assay was used to evaluate the ability of BIU-87 cells to migrate in vitro. Flow cytometry was used to detect the effect of sirolimus on the cell cycle of BIU-87 cells. Results The expression level of Raptor was significantly decreased, the migration ability of BIU-87 cells was significantly decreased in a dose-dependent manner, in sirolimus treated group than in the control. Sirolimus blocked BIU-87 cell cycle in G_0/G_1 phase, thereby inhibiting the cell proliferation. Conclusion The inhibition of Sirolimus on the proliferation and migration of BIU-87 cells might be related to inhibiting the expression of Raptor.
引文
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[11] Zhan Y, Shen LS, Xu WS, et al. Functional improvements in patients with lymphangioleiomyomatosis after sirolimus: an observational study[J]. Orphanet Journal of Rare Diseases, 2018, 20: 13(34): 102-109.
[2] Ma MKM, Yung S, Chan TM, et al. mTOR Inhibition and Kidney Diseases[J]. Transplantation, 2018, 102(2S Suppl 1): S32-S40.
[3] Baliu C, Esforzado N, Campistol JM, et al. Chronic lymphedema in renal transplant recipients under immunosuppression with sirolimus: presentation of 2 cases[J]. JAMA Dermatol, 2014, 150(9):1023-1024.
[4] Hara K, Maruki L, Long X, el a1. Raptor, a binding partner of target of rapamycin(mTOR), mediates TOR action[J]. Cell, 2002, 110(2): 177-189.
[5] Oshiro N, Yoshino K, Hidaya S, et al. Dissociation of raptor from mTOR is a mechanism of rapamycin-induced inhibition of mTOR function[J]. Genes Cell, 2004, 9(4): 359-366.
[6] Mussin N, Oh SC, Lee KW, et al. Sirolimus and Metformin Synergistically Inhibits Colon Cancer In Vitro and In Vivo[J]. J Korean Med Sci, 2017, 32(9): 1385-1395.
[7] Kawata T, Tada K, Kobayashi M, et al. Dual inhibition of the mTORC1 and mTORC2 signaling pathways is a promising therapeutic target for adult T-cell leukemia[J]. Cancer Sci, 2018,109(1): 103-111.
[8] Martin-Liberal J, López-Pousa A, Martínez-Trufero J, et al. Phase II Study of Gemcitabine Plus Sirolimus in Previously Treated Patients with Advanced Soft-Tissue Sarcoma: a Spanish Group for Research on Sarcomas (GEIS) Study[J]. Target Oncol, 2018, 13(1): 81-87.
[9] Wu KC, Huang HC, Chang T, et al. Effect of sirolimus on liver cirrhosis and hepatic encephalopathy of common bile duct-ligated rats[J]. Eur J Pharmacol, 2018, 824: 133-139.
[10] Korula S, Chapla A, Priyambada L, et al. Sirolimus therapy for congenital hyperinsulinism in an infant with a novel homozygous KCNJ11 mutation[J]. J Pediatr Endocrinol Metab, 2018, 31(1): 87-89.
[11] Zhan Y, Shen LS, Xu WS, et al. Functional improvements in patients with lymphangioleiomyomatosis after sirolimus: an observational study[J]. Orphanet Journal of Rare Diseases, 2018, 20: 13(34): 102-109.