4-1BB基因在不同分化胃癌细胞株中的表达

doi:10.3969/j.issn.1004-616x.2010.05.010

癌变·畸变·突变 ›› 2010, Vol. 22 ›› Issue (5): 370-373.doi: 10.3969/j.issn.1004-616x.2010.05.010

4-1BB基因在不同分化胃癌细胞株中的表达

宋振川1,王　磊1,郭宸君1,李　勇1,檀碧波2,范立侨1

1.河北医科大学第四医院外三科，河北石家庄 050011; 2. 河北省人民医院外二科，河北石家庄 050011

收稿日期:2010-03-29 修回日期:2010-06-02 出版日期:2010-09-30 发布日期:2010-09-30
通讯作者: 李　勇

Expression of 4-1BB in different gastric cancer cell lines

SONG Zhen-chuan1,WANG Lei1,GUO Chen-jun1,LI Yong1,,TAN Bi-bo2, FAN Li-qiao1

1. Third Department of Surgery, The Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei；2. Second Department of Surgery, The Peoples' Hospital of Hebei Province. Shijiazhuang 050011 Hebei, China

Received:2010-03-29 Revised:2010-06-02 Online:2010-09-30 Published:2010-09-30
Contact: LI Yong

摘要/Abstract

摘要： 目的：研究共刺激因子4_1BB在人胃高分化腺癌细胞株MKN_28、人胃中分化腺癌细胞株SGC_7901、人胃低分化腺癌细胞株BGC_823、人胃粘液腺癌细胞株MGC_803中的表达及其生物学意义。方法：用RT_PCR法测定胃癌细胞株中4_1BB mRNA表达；免疫细胞化学法测定胃癌细胞株中4_1BB蛋白表达；MTT法测定人淋巴细胞对胃癌细胞的体外杀伤活性；ELISA法检测人淋巴细胞与胃癌细胞共培养上清液中IL_2、IL_10、TNF_α的含量。结果： RT_PCR结果示4_1BB mRNA表达率从高到低依次为：MGC_803(77.30％)、BGC_823(71.68％)、SGC_7901(40.06％)、MKN_28(37.65％)。免疫细胞化学结果显示4_1BB着色程度由深到浅依次为：MGC_803、BGC_823、SGC_7901、MKN_28。MTT结果显示各时间段(24、48、72 h)不同效靶比(10∶1，20∶1，40，∶1)下，人淋巴细胞对胃癌细胞的体外杀伤活性由高到低的顺序为：MKN_28、SGC_7901、BGC_823、MGC_803。ELISA结果显示在各效靶细胞比下，人淋巴细胞与胃癌细胞共培养上清液中IL_2、 TNF_α的含量由高到低的顺序均为: MKN_28、SGC_7901、BGC_823、MGC_803。而IL_10的含量由高到低的顺序为: MGC_803、BGC_823、SGC_7901、 MKN_28。结论：人胃癌细胞株MKN_28、SGC_7901、BGC_823、MGC_803中4_1BB基因的mRNA 和蛋白均有表达，且胃癌细胞株的恶性程度越高其4_1BB表达水平越高；淋巴细胞对低表达4_1BB胃癌细胞的杀伤力较高；提示4_1BB可能通过抑制细胞因子 TNF_α、IL_2和促进细胞因子IL_10的产生调节肿瘤免疫。

关键词: 胃癌细胞株, 肿瘤免疫, 4-1BB

Abstract: OBJECTIVE: To study the expression of 4_1BB and its biological function in different human gastric cancer cell lines. METHODS: The expression of 4_1BB in different human gastric cancer cell lines was detected by immunohistochemical technique and reverse transcription polymerase chain reaction (RT_PCR). The killing activity of lymphocytes against gastric cancer cell lines was detected by MTT, whilst IL_2,IL_10,TNF_α were tested in the supernatant by ELISA. RESULTS: RT_PCR analysis showed that the 4_1BB mRNA expressions were MGC_803 (77.30％),BGC_823 (71.68％),SGC_7901(40.06％) and MKN_28(37.65％) in decreasing order. The 4_1BB protein expressions were MGC_803, BGC_823, SGC_7901 and MKN_28 in decreasing order by cytoimmunochemistry staining. MTT analysis showed that the killing activities from high to low were MKN_28, SGC_7901, BGC_823 and MGC_803. ELISA analysis showed that IL_2 and TNF_α levels were MKN_28, SGC_7901, BGC_823 and MGC_803 and IL_10 level was MGC_803,BGC_823, SGC_7901 and MKN_28, from high to low. CONCLUSION: The four different gastric cancer cell lines all expressed 4_1BB at mRNA and protein levels. The 4_1BB expression level related to gastric cancer cell lines to different degrees. The killing activity of lymphocytes against gastric cancer cell lines increased with decreasing 4_1BB expression. 4_1BB could regulate tumor immunity by inhibiting cell factors TNF_α and IL_2 and promoting cell factor IL_10.

Key words: gastric cancer cell lines, tumor immunity, 4-1BB

宋振川, 王　磊, 郭宸君, 李　勇, 檀碧波, 范立侨. 4-1BB基因在不同分化胃癌细胞株中的表达[J]. 癌变·畸变·突变, 2010, 22(5): 370-373.

SONG Zhen-chuan, WANG Lei, GUO Chen-jun, LI Yong, , TAN Bi-bo, FAN Li-qiao. Expression of 4-1BB in different gastric cancer cell lines[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2010, 22(5): 370-373.

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[2]	高　飞1/由　田2/吕晶玉3,/王运杰4. CAGE基因在颅脑肿瘤中的表达及其机制[J]. , 2007, 19(3): 239-242.
[3]	李巧霞单保恩 艾　军李　宏付小梅. 4-1BBL修饰小鼠结肠癌疫苗体外诱导抗肿瘤免疫反应的研究[J]. , 2007, 19(2): 103-105.

4-1BB基因在不同分化胃癌细胞株中的表达

Expression of 4-1BB in different gastric cancer cell lines

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