PDSW联合热疗对胃癌细胞和组织中COX-2、Bcl-2表达的影响
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摘要
旨在探讨生理性深层海水(PDSW)联合热疗对人胃癌SGC-7901细胞和组织中COX-2和Bcl-2表达的影响,明确其在人胃癌SGC-7901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,用实时荧光定量PCR检测细胞和组织中COX-2和Bcl-2基因表达情况,Western blot检测COX-2和Bcl-2蛋白的表达情况。结果显示,在SGC-7901细胞中和裸鼠皮下移植瘤的组织中,热疗均能降低COX-2、Bcl-2基因及蛋白的表达,同时,与生理盐水组相比,PDSW联合热疗处理下,细胞和组织中COX-2、Bcl-2表达量显著下降。PDSW联合热疗能抑制胃癌SGC-7901细胞中COX-2、Bcl-2基因和蛋白的表达,从而有效抑制胃癌细胞的增殖。
This work aims to explore the effect of PDSW combined with hyperthermia on COX-2 and Bcl-2 expressions in gastric cancercells SGC-7901 and tissues,and to clarify its anti-cancer effects on gastric cancer cells SGC-7901 in vitro and in vivo. The SGC-7901 cells werecultured in vitro and then were randomly divided into experimental,control and blank groups;the experimental group was added with PDSW,control group was given with normal saline,blank group was in no other treatment,and each group were cultured under 37℃,40℃ and 43℃respectively. Concurrently,the SGC-7901 cells were injected into the nude mice subcutaneous to establish the transplantation tumor model. Theinjected mice were randomly divided into 5 groups,and then given the treatment with H_2O+RT,H_2O +40℃,H_2O +43℃,PDSW+40℃,and PDSW+43℃ respectively when the tumor cells grew to 1 cm diameter. q RT-PCR and Western blot were used to detect the m RNA ofgene COX-2 and Bcl-2 and the protein expression of COX-2 and Bcl-2 in the cells and tissues. As results,in SGC-7901 cells and nude micesubcutaneous transplantation tumor tissues,the expressions of gene COX-2 and Bcl-2 m RNA and their protein expressions decreased underthe treatment of hyperthermia. Compared with the treatment of normal saline group,the expressions of gene COX-2 and Bcl-2 and their proteinexpression decreased remarkably under the treatment of PDSW combined with hyperthermia. Conclusively,PDSW combined with hyperthermiamay inhibit the expressions of gene COX-2 and Bcl-2 and their protein expressions in gastric cancer SGC-7901 cells and tissues,so as toeffectively inhibit the proliferation of gastric cancer cells.
This work aims to explore the effect of PDSW combined with hyperthermia on COX-2 and Bcl-2 expressions in gastric cancercells SGC-7901 and tissues,and to clarify its anti-cancer effects on gastric cancer cells SGC-7901 in vitro and in vivo. The SGC-7901 cells werecultured in vitro and then were randomly divided into experimental,control and blank groups;the experimental group was added with PDSW,control group was given with normal saline,blank group was in no other treatment,and each group were cultured under 37℃,40℃ and 43℃respectively. Concurrently,the SGC-7901 cells were injected into the nude mice subcutaneous to establish the transplantation tumor model. Theinjected mice were randomly divided into 5 groups,and then given the treatment with H_2O+RT,H_2O +40℃,H_2O +43℃,PDSW+40℃,and PDSW+43℃ respectively when the tumor cells grew to 1 cm diameter. q RT-PCR and Western blot were used to detect the m RNA ofgene COX-2 and Bcl-2 and the protein expression of COX-2 and Bcl-2 in the cells and tissues. As results,in SGC-7901 cells and nude micesubcutaneous transplantation tumor tissues,the expressions of gene COX-2 and Bcl-2 m RNA and their protein expressions decreased underthe treatment of hyperthermia. Compared with the treatment of normal saline group,the expressions of gene COX-2 and Bcl-2 and their proteinexpression decreased remarkably under the treatment of PDSW combined with hyperthermia. Conclusively,PDSW combined with hyperthermiamay inhibit the expressions of gene COX-2 and Bcl-2 and their protein expressions in gastric cancer SGC-7901 cells and tissues,so as toeffectively inhibit the proliferation of gastric cancer cells.
引文
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[15]Leung WK,To RF,Ng YP,et al.Association betweencyclooxygenase-2 overexpression and missense P53 mutations,ingastric cancer[J].Br J Cancer,2001,84(3):335-339.
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[17]李杰,杨红,郝洪岭,等.环氧合酶2与肿瘤研究进展[J].临床误诊误治,2012,25(4):73-75.
[18]陈美霓,郭浩,郭巍,等.COX-2蛋白和VEGF-C蛋白在胃癌和非胃癌黏膜病变组织中的表达及临床意义[J].海南医学,2015,26(18):2671-2674.
[19]Forones NM,Carvalho AP,Giannotti-Filho O,et al.Cellproliferation and apoptosis in gastric cancer and intestinalmetaplasia[J].Arq Gastroenterol,2005,42(1):30-34.
[20]Anagnostopoulos GK,Stefanou D,Arkoumani E,et al.Bax and Bcl-2 protein expression in gastric precancerous lesions:immunohistochemical study[J].J Gastroenterol Hepatol,2005,20(11):1674-1678.
[21]田明,张云锋,杜宁,等.PTEN和bcl-2在胃癌组织中的表达及临床意义[J].陕西医学杂志,2011,40(2):148-151.
[22]彭鹏,邹文蓉.p53,VEGF,Bcl-2在进展期胃癌组织中的表达[J].现代医药卫生,2011,27(2):184-186.
[2]张佳慧,秦丽娟.肿瘤热疗的研究进展[J].前沿进展,2012,9:1424-1426.
[3]闫向勇,刘文超.热疗在肿瘤治疗中的研究进展[J].世界中西医结合杂志,2014,9(2):213-216.
[4]崔进.“深层海水”在医学领域中的研究[J].昆明医学院学报,2011,32(8):l-2.
[5]Kozak KR,Rowlinson SW,Marnett LJ.Oxygenation of theendocannabinoid,2-arachidonylglycerol,to glyceryl prostaglandinsby cyclooxygenase-2[J].J Biol Chem,2000,275:33744-33749.
[6]Radmark O,Shimizu T,Jornvall H,et al.Leukotriene A4 hydrolasein human leukocytes.Purification and properties[J].J Biol Chem,1984,259:12339-12345.
[7]李士坤,任庆梅,陈克河.不同类型胃息肉组织中COX-2基因蛋白的表达及其意义[J].中国实用医药,2012,7(29):51-52.
[8]田慧军,邓涛,沈志祥,等.大肠癌和大肠腺瘤COX-2和Bcl-2的基因表达及其意义[J].胃肠病学和肝病学杂志,2003,12(6):544-546.
[9]肖炜明,施瑞华,丁岩冰,等.胃癌组EGFR和COX-2表达的意义及其相关性[J].世界华人消化杂志,2007,15(2):123.
[10]李为明,崔进,徐鹏远,代佑果,深层海水对小鼠创面愈合的促进作用[J].重庆医学,2014,2(43):462-464.
[11]Tang R,Zhu ZG,Qu Y,et al.The impact of hyperthermicchemotherapy on human gastric cancer cell lines:preliminaryresults[J].Oncol Rep 2006,16(3):549-559.
[12]代佑果.“深层海水”与热疗联合治疗肝细胞癌的实验性研究[M].昆明医科大学,2014,5.
[13]Miladi-Abdennadher I,Abdelmaksoud-Dammak R,Ayed-GuerfaliDB,et al.Expression of COX-2 and E-cadherin in Tunisian patientswith colorectal adenocarcinoma[J].Acta Histochemica,2012,114(6):577-581.
[14]Serra KP,Sarian LO,Rodrigues-Peres RM,et al.Expression ofcyclooxygenase-2(COX-2)and p53 in neighboring invasive andin situ components of breast tumors[J].Acta Histochemica,2012,114(3):226-231.
[15]Leung WK,To RF,Ng YP,et al.Association betweencyclooxygenase-2 overexpression and missense P53 mutations,ingastric cancer[J].Br J Cancer,2001,84(3):335-339.
[16]李百文,沈强.胃癌不同临床病理分期中环氧合酶与E基质金属蛋白酶的表达及其相关性[J].复旦学报:医学版,2007,34(1):223.
[17]李杰,杨红,郝洪岭,等.环氧合酶2与肿瘤研究进展[J].临床误诊误治,2012,25(4):73-75.
[18]陈美霓,郭浩,郭巍,等.COX-2蛋白和VEGF-C蛋白在胃癌和非胃癌黏膜病变组织中的表达及临床意义[J].海南医学,2015,26(18):2671-2674.
[19]Forones NM,Carvalho AP,Giannotti-Filho O,et al.Cellproliferation and apoptosis in gastric cancer and intestinalmetaplasia[J].Arq Gastroenterol,2005,42(1):30-34.
[20]Anagnostopoulos GK,Stefanou D,Arkoumani E,et al.Bax and Bcl-2 protein expression in gastric precancerous lesions:immunohistochemical study[J].J Gastroenterol Hepatol,2005,20(11):1674-1678.
[21]田明,张云锋,杜宁,等.PTEN和bcl-2在胃癌组织中的表达及临床意义[J].陕西医学杂志,2011,40(2):148-151.
[22]彭鹏,邹文蓉.p53,VEGF,Bcl-2在进展期胃癌组织中的表达[J].现代医药卫生,2011,27(2):184-186.