TRPC6在子宫内膜癌组织中的表达及对细胞增殖作用的研究
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摘要
目的探讨子宫内膜癌组织中瞬时受体电位阳离子通道-6(TRPC6)的表达变化及其在子宫内膜癌细胞增殖中的作用。方法选取2011年1月至2015年6月在河南省人民医院和许昌市人民医院手术治疗的子宫内膜癌患者30例作为研究组,24例不典型增生和28例子宫肌瘤患者作为对照组;应用分子生物学技术检测30例子宫内膜癌、24例不典型增生和28例正常子宫内膜组织中TRPC6的表达;用阻断剂SKF96365和siRNA干扰两种方法阻断TRPC6后,用细胞计数方法和氚胸腺嘧啶核苷掺入试验观察子宫内膜癌HEC-1A细胞增殖的变化。结果子宫内膜癌组织中的TRPC6 mRNA的表达量明显高于不典型增生组织和正常子宫内膜组织TRPC6 mRNA的表达量(0.98±0.56 vs. 0.30±0.24和0.23±0.13,P<0.01);子宫内膜癌组织中TRPC6蛋白的表达量明显高于不典型增生组织和正常子宫内膜组织中TRPC6蛋白的表达量(1.22±0.39 vs. 0.75±0.27和0.73±0.26,P<0.01);子宫内膜癌组织中TRPC6的表达水平与手术病理分期无关,但与病理分级有关;SKF96365呈剂量依赖性方式阻滞细胞增殖,使细胞生长曲线下移;转染siRNA细胞TRPC6蛋白表达量为转染阴性对照siRNA细胞的(38.51±6.21)%,与转染阴性对照相比,转染TRPC6 siRNA显著减少细胞的增殖。结论 TRPC6的异常表达与子宫内膜癌细胞的增殖相关,降低其表达可减少子宫内膜癌细胞的增殖。
Objective To study the expression of transient receptor potential canonical-6(TRPC6)channels in endometrial cancer tissues and their role in regulating proliferation of endometrial cancer cells.Methods Thirty patients with endometrial cancer who were treated from January 2011 to June 2015 in Henan Provincial People's Hospital and Xuchang People's Hospital were chosen as the study group;the controls group consisted of 24 atypical hyperplasia patients and 28 uterus leiomyoma patients.Molecular biological techniques were used to examine the expression of TRPC6 channels in 30 endometrial cancer specimens,24 atypical hyperplasia specimens and 28 normal endometrial specimens.SKF96365(an inhibitor of TRPC6 channel)and siRNA interference(RNAi)targeting TRPC6 channel were used to block TRPC6 so as to explore the role of TRPC6 channels in regulating the proliferation of endometrial cancer cells by[3 H]thymidine incorporation and cell number.Results The expression levels of TRPC6 in endometrial cancer were notably elevated than those in the atypical hyperplasia endometrial and normal endometrial tissues. The expression levels of TRPC6 in endometrial cancer vs. the control:mRNA:(0.98±0.56)vs.(0.30±0.24 and0.23±0.13)(P<0.01):protein:(1.22±0.39)vs.(0.75±0.27 and 0.73±0.26)(P<0.01);The expression level of TRPC6in endometrial cancer tissues was not related to the surgical pathological staging, but was related to pathological staging; SKF96365 caused a dosedependent decline in cell amount of HEC-1 A cell. The expression quantity of TRPC6 in whole lysates of the celltransfected with target-TRPC6 small interference RNA(siRNA)was(38.51±6.21)% of that found in the cells transfected with non-silencing RNA;[3 H]thymidine incorporation in HEC-1 A transfected with target-TRPC6 siRNA was also reduced,siRNA inhibited HEC-l A cells proliferation,compared with the cells transfected with non-silencing RNA.Conclusions TRPC6 channels mignt be closely related to the proliferation of endometrial cancer cells and down regulation of its expression may suppress its development.
Objective To study the expression of transient receptor potential canonical-6(TRPC6)channels in endometrial cancer tissues and their role in regulating proliferation of endometrial cancer cells.Methods Thirty patients with endometrial cancer who were treated from January 2011 to June 2015 in Henan Provincial People's Hospital and Xuchang People's Hospital were chosen as the study group;the controls group consisted of 24 atypical hyperplasia patients and 28 uterus leiomyoma patients.Molecular biological techniques were used to examine the expression of TRPC6 channels in 30 endometrial cancer specimens,24 atypical hyperplasia specimens and 28 normal endometrial specimens.SKF96365(an inhibitor of TRPC6 channel)and siRNA interference(RNAi)targeting TRPC6 channel were used to block TRPC6 so as to explore the role of TRPC6 channels in regulating the proliferation of endometrial cancer cells by[3 H]thymidine incorporation and cell number.Results The expression levels of TRPC6 in endometrial cancer were notably elevated than those in the atypical hyperplasia endometrial and normal endometrial tissues. The expression levels of TRPC6 in endometrial cancer vs. the control:mRNA:(0.98±0.56)vs.(0.30±0.24 and0.23±0.13)(P<0.01):protein:(1.22±0.39)vs.(0.75±0.27 and 0.73±0.26)(P<0.01);The expression level of TRPC6in endometrial cancer tissues was not related to the surgical pathological staging, but was related to pathological staging; SKF96365 caused a dosedependent decline in cell amount of HEC-1 A cell. The expression quantity of TRPC6 in whole lysates of the celltransfected with target-TRPC6 small interference RNA(siRNA)was(38.51±6.21)% of that found in the cells transfected with non-silencing RNA;[3 H]thymidine incorporation in HEC-1 A transfected with target-TRPC6 siRNA was also reduced,siRNA inhibited HEC-l A cells proliferation,compared with the cells transfected with non-silencing RNA.Conclusions TRPC6 channels mignt be closely related to the proliferation of endometrial cancer cells and down regulation of its expression may suppress its development.
引文
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[2]中国抗癌协会妇科肿瘤专业委员会.子宫内膜癌诊断与治疗指南(第四版)[J].中国实用妇科与产科杂志,2018,34(8):880-886.
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[6]Chigurupati S,Venkataraman R,Barrera D,et al.Receptor channel TRPC6 is a key mediator of Notch-driven glioblastoma growth and invasiveness[J].Cancer Res,2010,70(1):418-427.
[7]Ding X,He Z,Zhou K,et al.Essential role of TRPC6 channels in G2/M phase transition and development of human glioma[J].J Natl Cancer Inst,2010,102(14):1052-1068.
[8]Wan Q,Zheng A,Liu X,et al.Expression of transient receptor potential channel 6 in cervical cancer[J].Onco Targets Ther,2012,(5):171-176.
[9]Song J,Wang Y,Li X,et al.Critical role of TRPC6 channels in the development of human renal cell carcinoma[J].Mol Biol Rep,2013,40(8):5115-5122.
[10]Guilbert A,Dhennin-Duthille I,Hiani YE,et al.Expression of TRPC6 channels in human epithelial breast cancer cells[J].BMC Cancer,2008,8:125.
[11]Ding M,Wang H,Qu C,et al.Pyrazolo[1,5-a]pyrimidine TRPC6 antagonists for the treatment of gastric cancer[J].Cancer Lett,2018,432(28):47-55.
[12]Bernichtein S,Pigat N,Barry DN,et al.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.
[13]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.
[14]Isaac J,Diez-BelloR.TRPC6 Channels are required for proliferation,migration and invasion of breast cancer cell lines by modulation of orail and orail surface exposure[J].Cancers,2018,10(9):E331.
[15]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.
[16]Wang Y,Yue D,Li K,et al.The role of TRPC6 in HGF-induced cell proliferation of human prostate cancer DU145 and PC3 cells[J].Asian J Androl,2010,12(6):841-852.2018-12-152019-02-14
[2]中国抗癌协会妇科肿瘤专业委员会.子宫内膜癌诊断与治疗指南(第四版)[J].中国实用妇科与产科杂志,2018,34(8):880-886.
[3]宗昊,刘林枝,张岩.血浆瘦素水平与子宫内膜病变风险相关性研究[J].中国实用妇科与产科杂志,2018,34(6):662-665.
[4]王朕华,丰有吉,苏敏,等.钙激活性中电导钾离子通道在子宫内膜癌组织中的表达及其在细胞增殖中的作用[J].中华妇产科杂志,2007,42(2):111-115.
[5]ThebaμLt S,Flourakis M,Vanoverberghe K,et al.Differential role of transient receptor potential channels in Ca2+entry and proliferation of prostate cancer epithelial cells[J].Cancer Res,2006,66(4):2038-2047.
[6]Chigurupati S,Venkataraman R,Barrera D,et al.Receptor channel TRPC6 is a key mediator of Notch-driven glioblastoma growth and invasiveness[J].Cancer Res,2010,70(1):418-427.
[7]Ding X,He Z,Zhou K,et al.Essential role of TRPC6 channels in G2/M phase transition and development of human glioma[J].J Natl Cancer Inst,2010,102(14):1052-1068.
[8]Wan Q,Zheng A,Liu X,et al.Expression of transient receptor potential channel 6 in cervical cancer[J].Onco Targets Ther,2012,(5):171-176.
[9]Song J,Wang Y,Li X,et al.Critical role of TRPC6 channels in the development of human renal cell carcinoma[J].Mol Biol Rep,2013,40(8):5115-5122.
[10]Guilbert A,Dhennin-Duthille I,Hiani YE,et al.Expression of TRPC6 channels in human epithelial breast cancer cells[J].BMC Cancer,2008,8:125.
[11]Ding M,Wang H,Qu C,et al.Pyrazolo[1,5-a]pyrimidine TRPC6 antagonists for the treatment of gastric cancer[J].Cancer Lett,2018,432(28):47-55.
[12]Bernichtein S,Pigat N,Barry DN,et al.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.
[13]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.
[14]Isaac J,Diez-BelloR.TRPC6 Channels are required for proliferation,migration and invasion of breast cancer cell lines by modulation of orail and orail surface exposure[J].Cancers,2018,10(9):E331.
[15]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.
[16]Wang Y,Yue D,Li K,et al.The role of TRPC6 in HGF-induced cell proliferation of human prostate cancer DU145 and PC3 cells[J].Asian J Androl,2010,12(6):841-852.2018-12-152019-02-14