阻断TRPC6通道对人子宫内膜癌细胞和裸鼠移植瘤放射敏感性的影响
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摘要
目的:探讨阻断瞬时受体电位阳离子通道6(TRPC6)对子宫内膜癌细胞和裸鼠移植瘤放射敏感性的影响。方法:选用高表达TRPC6的子宫内膜癌细胞株HEC-1A,用阻断剂SKF96365阻断子宫内膜癌细胞中TRPC6表达,流式细胞仪检测细胞周期。将HEC-1A细胞分为4组:对照组、单纯SKF96365组、单纯照射组和SKF96365+照射组(10μmol/L SKF96365+放射),计算细胞克隆数,判断细胞的增殖能力和放射增敏率。将32只雌性裸鼠随机分为4组,每组8只,分别为单纯SKF96365组、单纯照射组、SKF96365+照射组和对照组,右腹皮下注射5×10~6个细胞,每周测量裸鼠皮下肿瘤体积,接种12周后处死小鼠,测肿瘤的重量和体积,计算肿瘤生长延迟时间(TGD)和放射增敏因子。结果:SKF96365处理后,子宫内膜癌HEC-1A细胞中G_2/M期细胞百分比明显增加,G_0/G_1期百分比减少。SKF96365+照射组的细胞克隆数最低。裸鼠的实验结果与细胞实验结果相似,SKF96365+照射组裸鼠的皮下移植瘤体积和重量最小。结论:阻断TRPC6通道对子宫内膜癌HEC-1A细胞和裸鼠的肿瘤均有明显的放射增敏作用。TRPC6通道可作为子宫内膜癌放射增敏的调控点,可能成为子宫内膜癌治疗的新靶点。
Objective:To investigate the effect of blockage of transient receptor potential-6(TRPC6)channel on the radiation sensitization of endometrial carcinoma cells and cancer xenograft in nude mice.Methods:SKF96365(an inhibitor of TRPC6 channel) was used to block TRPC6 so as to explore the role of TRPC6 channels in regulating the cell cycle of endometrial cancer cells by flow cytometry.HEC-1 A cells were treated with radiation after 12 hours of incubation of 10μmol/L SKF96365,and the cell clones were counted and the proliferation of the treated cells were estimated and sensitive enhancement ratio(SER) was figured up.Thirty-two female nude mice were randomly divided into 4 groups,8 in each group:control group,SKF963365 alone group,irradiation alone group,and SKF96365+ irradiation group.5×10~6 cells were injected subcutaneously into the right flank regions of each mice.Mice were checked everyweek for tumor appearance,and the volume of tumor were measured.Twelve weeks after inoculation,all mice were killed by cervical dislocation,and the volume and weight of tumor were got.Tumor growth delay time and enhancement factor were figured up. Results: The percentage of G2/M phase cells in HEC-1 A cells treated with SKF96365 was significantly increased,and the percentage of cells in G0/G1 was decreased,cell clone number( the number of cells was greater than 100) that received SKF96365 + irradiation were less than the others treatments. In vivo studies showed similar results to those of the above-mentioned cell experiments,and the nude mice of the SKF96365 + irradiation group had the less volume and weight of subcutaneously transplanted tumors than others groups.Conclusion: Blocking the TRPC6 channel has a significant effect on the radiation sensitization in both endometrial carcinoma HEC-1 A cells and cancer xenograft in nude mice. TRPC6 channel can be used as a regulation point of endometrial cancer radiosensitization,and it has potential to be a new therapeutic target for endometrial carcinoma.
Objective:To investigate the effect of blockage of transient receptor potential-6(TRPC6)channel on the radiation sensitization of endometrial carcinoma cells and cancer xenograft in nude mice.Methods:SKF96365(an inhibitor of TRPC6 channel) was used to block TRPC6 so as to explore the role of TRPC6 channels in regulating the cell cycle of endometrial cancer cells by flow cytometry.HEC-1 A cells were treated with radiation after 12 hours of incubation of 10μmol/L SKF96365,and the cell clones were counted and the proliferation of the treated cells were estimated and sensitive enhancement ratio(SER) was figured up.Thirty-two female nude mice were randomly divided into 4 groups,8 in each group:control group,SKF963365 alone group,irradiation alone group,and SKF96365+ irradiation group.5×10~6 cells were injected subcutaneously into the right flank regions of each mice.Mice were checked everyweek for tumor appearance,and the volume of tumor were measured.Twelve weeks after inoculation,all mice were killed by cervical dislocation,and the volume and weight of tumor were got.Tumor growth delay time and enhancement factor were figured up. Results: The percentage of G2/M phase cells in HEC-1 A cells treated with SKF96365 was significantly increased,and the percentage of cells in G0/G1 was decreased,cell clone number( the number of cells was greater than 100) that received SKF96365 + irradiation were less than the others treatments. In vivo studies showed similar results to those of the above-mentioned cell experiments,and the nude mice of the SKF96365 + irradiation group had the less volume and weight of subcutaneously transplanted tumors than others groups.Conclusion: Blocking the TRPC6 channel has a significant effect on the radiation sensitization in both endometrial carcinoma HEC-1 A cells and cancer xenograft in nude mice. TRPC6 channel can be used as a regulation point of endometrial cancer radiosensitization,and it has potential to be a new therapeutic target for endometrial carcinoma.
引文
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[12] Sozucan Y,Kalender ME,Sari I,et al.TRP genes family expression in colorectal cancer[J].Exp Oncol,2015,37(3):208-212
[13] Zeng B,Yuan C,Yang X,et al.TRPC channels and their splice variants are essential for promoting human ovarian cancer cell proliferation and tumorigenesis[J].Curr Cancer Drug Targets,2013,13(1):103-116
[14] Wu G,Chen G,Zhou J,et al.Liriodenine enhances radiosensitivity in esophageal cancer ECA-109 cells by inducing apoptosis and G2/M arrest[J].Oncol Lett,2018,16(4):5020-5026
[15] Storch K,Dickreuter E,Artati A,et al.BEMER electromagnetic field therapy reduces cancer cell radioresistance by enhanced ROS formation and induced DNA damage[J].PLoS One,2016,11(12):e0167931
[16] Assani G,Zhou Y.Effect of modulation of epithelial-mesenchymal transition regulators Snail1 and Snail2 on cancer cell radiosensitivity by targeting of the cell cycle,cell apoptosis and cell migration/invasion[J].Oncol Lett,2019,17(1),23-30
[17] Zheng R,Liu Y,Zhang X,et al.miRNA-200c enhances radiosensitivity of esophageal cancer by cell cycle arrest and targeting P21[J].Biomed Pharmacother,2017,90:517-523
[2] Ozen A,Falchook AD,Varia MA,et al.Effect of race and histology on patterns of failure in women with early stage endometrial cancer treated with high dose rate brachytherapy[J].Gynecol Oncol,2015,138(2):429-433
[3] Harkenrider MM,Block AM,Siddiqui ZA,et al.The role of vaginal cuff brachytherapy in endometrial cancer[J].Gynecol Oncol,2015,136(2):365-372
[4] Delishaj D,Barcellini A,D'Amico R,et al.Vaginal toxicity after high-dose-rate endovaginal brachytherapy:20 years of results[J].J Contemp Brachytherapy,2018,10(6):559-566
[5] Siegel RL,Miller KD,Jemal A.Cancer Statistics,2017[J].CA Cancer J Clin,2017,67(1):7-30
[6] Pinto MC,Kihara AH,Goulart VA.Calcium signaling and cell proliferation[J].Cell Signal,2015,27(11):2139-2149
[7] Lee JM,Davis FM,Roberts-Thomson SJ,et al.Ion channels and transporters in cancer.4.Remodeling of Ca(2+) signaling in tumorigenesis:role of Ca2+transport[J].Am J Physiol Cell Physiol,2011,301(5):C969-976
[8] Xu J,Yang Y,Xie R,et al.The NCX1/TRPC6 complex mediates TGF β-Driven migration and invasion of human hepatocellular carcinoma cells[J].Cancer Res,2018,78(10):2564-2576
[9] Ding M,Wang H,Qu C,et al.Pyrazolo[1,5-a]pyrimidine TRPC6 antagonists for the treatment of gastric cancer[J].Cancer Lett,2018,28(432):47-55
[10] Yang LL,Liu BC,Lu XY,et al.Inhibition of TRPC6 reduces non-small cell lung cancer cell proliferation and invasion[J].Oncotarget,2017,8(3):5123-5134
[11] Bernichtein S,Pigat N,Barry Delongchamps,et al.N Vitamin D3 prevents calcium-induced progression of early-stage prostate tumors by counteracting trpc6 and calcium sensing receptor upregulation[J].Cancer Res,2017,77(2):355-365
[12] Sozucan Y,Kalender ME,Sari I,et al.TRP genes family expression in colorectal cancer[J].Exp Oncol,2015,37(3):208-212
[13] Zeng B,Yuan C,Yang X,et al.TRPC channels and their splice variants are essential for promoting human ovarian cancer cell proliferation and tumorigenesis[J].Curr Cancer Drug Targets,2013,13(1):103-116
[14] Wu G,Chen G,Zhou J,et al.Liriodenine enhances radiosensitivity in esophageal cancer ECA-109 cells by inducing apoptosis and G2/M arrest[J].Oncol Lett,2018,16(4):5020-5026
[15] Storch K,Dickreuter E,Artati A,et al.BEMER electromagnetic field therapy reduces cancer cell radioresistance by enhanced ROS formation and induced DNA damage[J].PLoS One,2016,11(12):e0167931
[16] Assani G,Zhou Y.Effect of modulation of epithelial-mesenchymal transition regulators Snail1 and Snail2 on cancer cell radiosensitivity by targeting of the cell cycle,cell apoptosis and cell migration/invasion[J].Oncol Lett,2019,17(1),23-30
[17] Zheng R,Liu Y,Zhang X,et al.miRNA-200c enhances radiosensitivity of esophageal cancer by cell cycle arrest and targeting P21[J].Biomed Pharmacother,2017,90:517-523
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