PDSW联合热疗可抑制胃癌NF-κB的表达
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摘要
为探讨生理性深层海水(PDSW)联合热疗对人胃癌SGC-7901细胞体外和体内的抗癌作用,通过在体外培养人胃癌SGC-7901细胞株,随机分为实验组、对照组和空白组,实验组加PDSW,对照组加生理盐水,空白组不做其他处理,分别在37℃、40℃和43℃热疗温度下培养。同时,将SGC-7901细胞注射至裸鼠皮下,建立移植瘤模型,随机分为5组,至肿瘤长至直径为1 cm时,分别进行H_2O+RT、H_2O+40℃、H_2O+43℃、PDSW+40℃、PDSW+43℃处理,然后绘制裸鼠肿瘤生长曲线,观察裸鼠肿瘤组织切片,并用实时荧光定量PCR检测细胞和组织中NF-κB基因表达情况,Western blotting检测NF-κB蛋白的表达情况。研究发现,与其他处理相比,PDSW联合热疗处理后肿瘤生长缓慢,肿瘤细胞的增长明显受抑,细胞和裸鼠皮下移植瘤组织中NF-κB基因和蛋白的表达量较低。说明PDSW联合热疗能抑制胃癌SGC-7901细胞中NF-κB基因和蛋白的表达,从而有效抑制胃癌细胞的增殖。
In order to explore the anti-cancer effects of PDSW combined with hyperthermia on gastric cancer cells SGC-7901 in vitro and in vitro, the SGC-7901 cells were cultured in vitro and then were randomly divided into experimental, control and blank groups. The experimental group was treated with PDSW, control group was treated with normal saline, and blank group had no other treatment. Each group was cultured respectively under37℃, 40℃and 43℃. At the same time, the SGC-7901 cells were injected into the nude mice to establish the model of transplantation tumor. Then they were divided into 5 groups randomly to treat with H_2 O+RT, H_2 O+40℃,H_2 O +43℃, PDSW+40℃, PDSW+43℃ when the tumour cells grew to a diameter of 1 cm. Then the growth curve of nude mice tumour was drawn, and tumor tissue sections were observed. Real time fluorescent quantitative PCR was used to detect the expression of NF-κB gene in cells and tissues, and Western blotting was used to detect the expression of NF-κB protein. It showed that compared with other treatments, the tumors grew slower and were significantly inhibited under the treatment of PDSW combined with hyperthermia. Also, the expression of NF-κB gene and protein were lower in the SGC-7901 cell and subcutaneous transplantation tumor tissue of nude mice.The study indicated that the PDSW combined with hyperthermia could inhibit the expression of NF-κB gene and protein, which could effectively inhibit the proliferation of gastric cancer cells.
In order to explore the anti-cancer effects of PDSW combined with hyperthermia on gastric cancer cells SGC-7901 in vitro and in vitro, the SGC-7901 cells were cultured in vitro and then were randomly divided into experimental, control and blank groups. The experimental group was treated with PDSW, control group was treated with normal saline, and blank group had no other treatment. Each group was cultured respectively under37℃, 40℃and 43℃. At the same time, the SGC-7901 cells were injected into the nude mice to establish the model of transplantation tumor. Then they were divided into 5 groups randomly to treat with H_2 O+RT, H_2 O+40℃,H_2 O +43℃, PDSW+40℃, PDSW+43℃ when the tumour cells grew to a diameter of 1 cm. Then the growth curve of nude mice tumour was drawn, and tumor tissue sections were observed. Real time fluorescent quantitative PCR was used to detect the expression of NF-κB gene in cells and tissues, and Western blotting was used to detect the expression of NF-κB protein. It showed that compared with other treatments, the tumors grew slower and were significantly inhibited under the treatment of PDSW combined with hyperthermia. Also, the expression of NF-κB gene and protein were lower in the SGC-7901 cell and subcutaneous transplantation tumor tissue of nude mice.The study indicated that the PDSW combined with hyperthermia could inhibit the expression of NF-κB gene and protein, which could effectively inhibit the proliferation of gastric cancer cells.
引文
Chen Y.N.,Cao J.,Su J.J.,Li Y.,and Zhu Z.Y.,2006,Expression and significance of Rel m RNA and NF-κB2 m RNA in hepatocellular carcinoma,Zhongshan Daxue Xuebao(Journal of Sun Yat-Sen University(Medical Sciences Edition)),27(6):677-680(陈远能,曹骥,苏建家,李瑗,朱振宇,2006,Rel m RNA和NF-κB m RNA在人肝细胞癌中的表达及其临床意义,中山大学学报(医学科学版),27(6):677-680)
Dai Y.G.,2014,PDSW combined with whole body hyperthermia treats hepatocellular carcinoma in mice,Dissertation for Ph.D.,Kunming Medical University,Supervisor:Cui J.,pp.5(代佑果,2014,“深层海水”与热疗联合治疗肝细胞癌的实验性研究,博士学位论文,昆明医科大学,导师:崔进,pp.5)
Li W.M.,Cui J.,Xu P.Y.,and Dai Y.G.,2014,The influence of deep sea water on wound healing of mice,Chongqing Yixue(Chongqing Medical Science),43(4):462-464(李为明,崔进,徐鹏远,代佑果,2014,深层海水对小鼠创面愈合的促进作用,重庆医学,43(4):462-464)
Sethi G.,Sung B.,and Aggarwal B.B.,2008,Nuclear factor-kappa B activation:from bench to bedside,Experimental Biology and Medicine,233(1):21-31
Shibata A.,Nagaya T.,Imai T.,Funahashi H.,Nakao A.,and Seo H.,2002,Inhibition of NF-κB activity decrease the VEGFm RNA expression in MDA-MB-231 breast cancer cells Breast Cancer Research and Treatment,73(3):237-243
Sun S.C.,and Ley S.C.,2008,New insights in to NF-κB regulation and function,Trends in Immunology,29(10):469-478
Tang H.R.,Cui J.,Zhang J.H.,Dai Y.G.,Li W.M.,and Liao C.2016,Effects of PDSW combined with hyperthermia treats on PTEN,Ki67 genes and protein expression of Gastric Cancer Cells SGC-7901,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(1):21-27(唐辉蓉,崔进,张家骅,代佑果,李为明,廖陈,2016,PDSW联合热疗对SGC-7901细胞PTEN和Ki67基因及蛋白表达的影响,基因组学与应用生物学,35(1):21-27)
Tang R.,Zhu Z.G.,Qu Y.,Li J.F.,Ji Y.B.,Cai Q.,and Lin Y.Z.2006,The impact of hyperthermic chemotherapy on human gastric cancer cell lines:preliminary results,Oncology Reports,16(3):631-641
Wang K.W.,Cao Q.H.,Lin Y.,Xiao P.,Che L.H.,Liao C.J.,and Xue L.,2010,Expression of NF-κB and IL-1ra in gastric cancer and its clinical significance,Zhongshan Daxue Xuebao(Journal Of Sun Yat-Sen University(Medical Sciences Edition)),31(1):95-99(王康玮,曹清华,林原,肖萍,车丽洪,罗灿峤,薛玲,2010,NF-κB及IL-1ra在胃癌中的表达及其意义,中山大学学报(医学科学版),31(1):95-99)
Yang N.,2014,Expression of NF-κB and Survivin in gastric carcinoma and its correlation,Thesis for M.S.,Yan'an University,Supervisor:Feng Y.C.,pp.6(杨宁,2014,NF-κB与Survivin在胃癌中的表达及其相关性研究,硕士学位论文,延安大学,导师:冯义朝,pp.6)
Zhang J.H.,and Qin L.J.,2012,Research progress of tumor hyperthermia department of physiology,Shiyong Xinnaofei Xueguanbing Zazhi(Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease),20(9):1424-1426(张佳慧和秦丽娟,2012,肿瘤热疗的研究进展,实用心脑肺血管病杂志,20(9):1424-1426)
Zhao N.,Zhou M.K.,Ye Y.W.,Chen Z.F.,Li Q.,and Zhou Y.N.,2012,Expression and significance of Notch1,Jagged1 and NF-κB in human gastric carcinoma,Nanjing Yike Daxue Xuebao(Acta Uniwersitatis Medicinalis Nanjing(Natural Science Edition)),32(8):1061-1065(赵娜,周梅恺,叶玉伟,陈兆峰,李强,周永宁,2012,胃癌组织中Notch1、Jagged1及NF-κB的表达及临床意义,南京医科大学学报(自然科学版),32(8):1061-1065)
Dai Y.G.,2014,PDSW combined with whole body hyperthermia treats hepatocellular carcinoma in mice,Dissertation for Ph.D.,Kunming Medical University,Supervisor:Cui J.,pp.5(代佑果,2014,“深层海水”与热疗联合治疗肝细胞癌的实验性研究,博士学位论文,昆明医科大学,导师:崔进,pp.5)
Li W.M.,Cui J.,Xu P.Y.,and Dai Y.G.,2014,The influence of deep sea water on wound healing of mice,Chongqing Yixue(Chongqing Medical Science),43(4):462-464(李为明,崔进,徐鹏远,代佑果,2014,深层海水对小鼠创面愈合的促进作用,重庆医学,43(4):462-464)
Sethi G.,Sung B.,and Aggarwal B.B.,2008,Nuclear factor-kappa B activation:from bench to bedside,Experimental Biology and Medicine,233(1):21-31
Shibata A.,Nagaya T.,Imai T.,Funahashi H.,Nakao A.,and Seo H.,2002,Inhibition of NF-κB activity decrease the VEGFm RNA expression in MDA-MB-231 breast cancer cells Breast Cancer Research and Treatment,73(3):237-243
Sun S.C.,and Ley S.C.,2008,New insights in to NF-κB regulation and function,Trends in Immunology,29(10):469-478
Tang H.R.,Cui J.,Zhang J.H.,Dai Y.G.,Li W.M.,and Liao C.2016,Effects of PDSW combined with hyperthermia treats on PTEN,Ki67 genes and protein expression of Gastric Cancer Cells SGC-7901,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(1):21-27(唐辉蓉,崔进,张家骅,代佑果,李为明,廖陈,2016,PDSW联合热疗对SGC-7901细胞PTEN和Ki67基因及蛋白表达的影响,基因组学与应用生物学,35(1):21-27)
Tang R.,Zhu Z.G.,Qu Y.,Li J.F.,Ji Y.B.,Cai Q.,and Lin Y.Z.2006,The impact of hyperthermic chemotherapy on human gastric cancer cell lines:preliminary results,Oncology Reports,16(3):631-641
Wang K.W.,Cao Q.H.,Lin Y.,Xiao P.,Che L.H.,Liao C.J.,and Xue L.,2010,Expression of NF-κB and IL-1ra in gastric cancer and its clinical significance,Zhongshan Daxue Xuebao(Journal Of Sun Yat-Sen University(Medical Sciences Edition)),31(1):95-99(王康玮,曹清华,林原,肖萍,车丽洪,罗灿峤,薛玲,2010,NF-κB及IL-1ra在胃癌中的表达及其意义,中山大学学报(医学科学版),31(1):95-99)
Yang N.,2014,Expression of NF-κB and Survivin in gastric carcinoma and its correlation,Thesis for M.S.,Yan'an University,Supervisor:Feng Y.C.,pp.6(杨宁,2014,NF-κB与Survivin在胃癌中的表达及其相关性研究,硕士学位论文,延安大学,导师:冯义朝,pp.6)
Zhang J.H.,and Qin L.J.,2012,Research progress of tumor hyperthermia department of physiology,Shiyong Xinnaofei Xueguanbing Zazhi(Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease),20(9):1424-1426(张佳慧和秦丽娟,2012,肿瘤热疗的研究进展,实用心脑肺血管病杂志,20(9):1424-1426)
Zhao N.,Zhou M.K.,Ye Y.W.,Chen Z.F.,Li Q.,and Zhou Y.N.,2012,Expression and significance of Notch1,Jagged1 and NF-κB in human gastric carcinoma,Nanjing Yike Daxue Xuebao(Acta Uniwersitatis Medicinalis Nanjing(Natural Science Edition)),32(8):1061-1065(赵娜,周梅恺,叶玉伟,陈兆峰,李强,周永宁,2012,胃癌组织中Notch1、Jagged1及NF-κB的表达及临床意义,南京医科大学学报(自然科学版),32(8):1061-1065)