凋亡大肠癌CT26细胞对小鼠T淋巴细胞增殖及活性的影响  

Effect of apoptotic colorectal cancer CT26 cells on proliferation and activity of T lymphocytes in mice

作　　者：孙朝文[1,2] 张广钰[1] 赵辰[1,2] 成怀福 钟漓[1] 戴凌[1] 

机构地区：[1]桂林医学院附属医院胃肠外科,桂林市541001 [2]桂林医学院研究生学院,桂林市541004

出　　处：《广西医学》2016年第9期1201-1206,1210,共7页Guangxi Medical Journal

基　　金：广西自然科学基金(2012GXNSFAA276035);广西桂林市科学研究与技术开发计划(20120121-1-13)

摘　　要：目的探讨凋亡大肠癌CT26细胞对小鼠T淋巴细胞增殖及活性的影响。方法 (1)取对数生长期大肠癌CT26细胞制备凋亡肿瘤细胞及坏死肿瘤细胞,取Balb/c小鼠脾脏分离T淋巴细胞,并制备细胞毒性T淋巴细胞(CTL)。(2)将凋亡肿瘤细胞、坏死肿瘤细胞、正常肿瘤细胞分别与小鼠CTL共同培养(分别为凋亡肿瘤细胞组、坏死肿瘤细胞组、肿瘤细胞对照组),测定CTL的活性及增殖情况。(3)将小鼠T淋巴细胞分为对照组、刀豆蛋白A(Con A)组、凋亡肿瘤细胞+Con A组、坏死肿瘤细胞+Con A组、活肿瘤细胞+Con A组,经相应处理后测定各组T淋巴细胞的活性及增殖情况。(4)选取30只Balb/c小鼠分为凋亡肿瘤细胞组、坏死肿瘤细胞组、肿瘤细胞对照组各10只,分别将凋亡、坏死、对数生长期CT26细胞通过皮下及静脉输注小鼠,检测各组小鼠血清调节性T细胞(Treg)的表达情况。(5)另取30只小鼠,按方法(4)分组及处理后,测定各组小鼠血清可溶性Fas(s Fas)、白细胞介素(IL)-12、干扰素-γ(IFN-γ)、IL-4、IL-10、转化生长因子-β(TGF-β)的表达水平。结果凋亡肿瘤细胞组中CTL的活性、A450值均低于其他两组(P<0.05)。凋亡肿瘤细胞+Con A组的T淋巴细胞CD69、CD25、CD71的表达率以及G2、G3及G4期的T淋巴细胞数量均低于Con A组、活肿瘤细胞+Con A及坏死肿瘤细胞+Con A(P<0.05)。凋亡肿瘤细胞组Treg的表达水平、s Fas相对表达量低于肿瘤细胞组、坏死肿瘤细胞组(P<0.05)。凋亡肿瘤细胞组IL-10、TGF-β表达水平高于其他两组(P<0.05),而IL-12表达水平均低于其他两组(P<0.05),IL-4表达水平则高于肿瘤细胞对照组(P<0.05)。结论凋亡小鼠大肠癌CT26细胞能够抑制小鼠T淋巴细胞增殖及活性,影响大肠癌小鼠正常免疫功能。Objective To investigate the effect of apoptotic colorectal cancer CT26 cells on the proliferation and activity of T lymphocytes in mice. Methods ① Colorectal cancer CT26 cells during logarithmic growth phase were used to prepare apoptotic tumor cells and necrotic tumor cells. T lymphocytes were isolated from the spleens of Balb/c mice, and then cytotoxic T lymphocytes （CTL） were prepared. ②The apoptotic tumor cells, necrotic tumor cells and normal tumor cells were cultured with murine CTL separately （ taken as apoptotic tumor cell group, necrotic tumor cell group and tumor cell control group, respectively）. Then the activity and proliferation of CTL were detected. ③ The T lymphoeytes of mice were divided into control group, concanavalin A（ConA） group, apoptotic tumor cell ＋ ConA group, necrotic tumor cell ＋ ConA group and living tumor cell ＋ ConA group. The activity and proliferation of T lymphocyte in each group were detected after corresponding treatment.④Thirty Balb/c mice were selected and divided into apoptotic tumor cell group, necrotic tumor cell group and tumor cell control group ,with 10 mice in each group. Apoptotic CT26 cells, necrotic CT26 cells and CT26 cells during logarithmic growth phase were subcutaneously and intravenously infused into mice in corresponding groups. Then the expression of regulatory T cells （Treg） in each group was detected. ⑤ Another 30 mice were selected,then were grouped and treated by the way of method ④. The expression levels of soluble Fas （ sFas ） , interleukin （ IL ） -12, interferon gamma （ IFN-γ ） , IL-4, IL-10 and transforming growth factor beta （TGF-β） in each group were detected. Results The activity and A450 value of CTL in the apoptotic tumor cell group were lower than those in the other two groups （P 〈 0.05 ）. The expression rates of T lymphocytes （including CD69, CD25 and CD71 ） and number of Tlymphocytes during stage G2 or G3 in the apoptotic tumor cell ＋ ConA group were lower than those in the ConA

关 键 词：大肠癌 CT26细胞 T淋巴细胞 活性 增殖 调节性T细胞 可溶性FAS 小鼠 

分 类 号：R735.3[医药卫生—肿瘤] R392.4[医药卫生—临床医学]