AIM To study the tumorigenicity of colorectal cancer cells transfected with B7 gene and the anti-tumor immunity induced by B7 gene modified colorectal cancer cells.METHODS B7 gene was transfected into mouse colon canc...AIM To study the tumorigenicity of colorectal cancer cells transfected with B7 gene and the anti-tumor immunity induced by B7 gene modified colorectal cancer cells.METHODS B7 gene was transfected into mouse colon cancer cell line CMT93. The transfectants were selected in DMEM containing 800 mg/ L G418, and B7 molecules were detected by immunohistochemistry. Experiments in vivo include: ① 5×106 B7+ CMT93 cells were inoculated into the back of C57BL/ 6 mice subcutaneously to determine their tumorigenicity (n=4). As control, wild type CMT93 cells were inoculated the same as the experimental group (n=3). ② The mice primed by B7+ CMT93 cells whose tumors vanished were rechallenged with wild type CMT93 to observe the immune protection of these mice against the wild type CMT93 (n=4). Non-primed 4 native mice inoculated with wild type CMT93 were used as control. With in vitro cytotoxicity assay, the mice were immunized with B7+ CMT93 or the wild type CMT93 by intraperitoneal injection (n=4×2). The spleen cells and the abdominal cavity infiltrating lymphocytes were obtained and cultured for two days. Cytotoxicity of these cells against the B7 gene modified or wild type CMT93 was detected by MTT assay.RESULTS B7 high expression clones were obtained after the transfection of the B7 gene into CMT93 cells by electroporation. Immunohistochemistry results showed mainly membrance staining and partly cytoplasm staining in B7 gene transfected CMT93 cells. In vivo experiments: ① After the inoculation of the B7+ CMT93 cells into the back of C57BL/ 6 mice, they lost their tumorigenicity greatly (P＜0.01). All the small tumors growing in the early period in the experimental group vanished in one month, and the tumors in control group grew progressively. ② No tumors were found in all 4 mice primed by B7+ CMT93 cells after they were rechallenged with wild type CMT93. In the control group all mice had grown tumors (P＜0.05). In vitro cytotoxicity assay, the CTLs induced by B7+ CMT93 had a higher cytotoxity against the wild type CMT93 than that induced by wild type CMT93 (P＜0.05), and the cytotoxity of CTLs induced by B7+ CMT93 against B7+ CMT93 cells was higher than that against wild type CMT93 cells (P＜0.05).CONCLUSION The results suggest that the expression of costimulation B7 molecules by colorectal cancer cells can decrease their tumorigenicity greatly, and the B7 molecule can augment the activation of the CTLs against colorectal cancer, and it plays an important role in CTL effector function as well.展开更多
背景与目的:EB病毒(Epstein-Barr virus,EBV)感染在鼻咽癌发生中发挥重要作用。本研究的目的在于探讨潜伏的EBV活化后形成的复发感染对鼻咽癌发生过程中全基因组表达的影响。方法:收集EBV阳性(EBV+)/EBV阴性(EBV-)的鼻咽癌靶标基因表达...背景与目的:EB病毒(Epstein-Barr virus,EBV)感染在鼻咽癌发生中发挥重要作用。本研究的目的在于探讨潜伏的EBV活化后形成的复发感染对鼻咽癌发生过程中全基因组表达的影响。方法:收集EBV阳性(EBV+)/EBV阴性(EBV-)的鼻咽癌靶标基因表达谱和EBV复发感染次数不同的鼻咽癌细胞株基因表达谱,通过生物信息学手段对其进行交叉比较,统计学分析筛选出25个差异基因。运用DAVID(database for annotation,visualization and integrated discovery)、pSTIING(protein,signaling,transcriptional interactions and inflammation networks gateway)、GATHER(gene annotation tool to help explain relationships)和TELiS(transcriptionelement listening system)在线分析工具对25个共同差异基因进行表达模式的预测。结果:与EBV初次感染时的基因表达谱比较,仅有DUSP1、TOP1、HOXA9、DEK、IMPDH2、PABPC1等25个基因在EBV复发感染时仍然呈明显差异表达;这25个差异基因及其相关转录因子主要通过2种模式相互作用:一条主要涉及TOP1、DUSP1、DUSP6和RPS28等,另一条是由PITX1、CD9、HOXA9和IMPDH2组成的转录相关环。结论:EBV潜伏后的复发感染,可能通过筛选到的差异基因的2种表达模式发挥功能,最终导致鼻咽癌的发生。展开更多
基金Supported by the National Natural Science Foundation of China,No.3950076.
文摘AIM To study the tumorigenicity of colorectal cancer cells transfected with B7 gene and the anti-tumor immunity induced by B7 gene modified colorectal cancer cells.METHODS B7 gene was transfected into mouse colon cancer cell line CMT93. The transfectants were selected in DMEM containing 800 mg/ L G418, and B7 molecules were detected by immunohistochemistry. Experiments in vivo include: ① 5×106 B7+ CMT93 cells were inoculated into the back of C57BL/ 6 mice subcutaneously to determine their tumorigenicity (n=4). As control, wild type CMT93 cells were inoculated the same as the experimental group (n=3). ② The mice primed by B7+ CMT93 cells whose tumors vanished were rechallenged with wild type CMT93 to observe the immune protection of these mice against the wild type CMT93 (n=4). Non-primed 4 native mice inoculated with wild type CMT93 were used as control. With in vitro cytotoxicity assay, the mice were immunized with B7+ CMT93 or the wild type CMT93 by intraperitoneal injection (n=4×2). The spleen cells and the abdominal cavity infiltrating lymphocytes were obtained and cultured for two days. Cytotoxicity of these cells against the B7 gene modified or wild type CMT93 was detected by MTT assay.RESULTS B7 high expression clones were obtained after the transfection of the B7 gene into CMT93 cells by electroporation. Immunohistochemistry results showed mainly membrance staining and partly cytoplasm staining in B7 gene transfected CMT93 cells. In vivo experiments: ① After the inoculation of the B7+ CMT93 cells into the back of C57BL/ 6 mice, they lost their tumorigenicity greatly (P＜0.01). All the small tumors growing in the early period in the experimental group vanished in one month, and the tumors in control group grew progressively. ② No tumors were found in all 4 mice primed by B7+ CMT93 cells after they were rechallenged with wild type CMT93. In the control group all mice had grown tumors (P＜0.05). In vitro cytotoxicity assay, the CTLs induced by B7+ CMT93 had a higher cytotoxity against the wild type CMT93 than that induced by wild type CMT93 (P＜0.05), and the cytotoxity of CTLs induced by B7+ CMT93 against B7+ CMT93 cells was higher than that against wild type CMT93 cells (P＜0.05).CONCLUSION The results suggest that the expression of costimulation B7 molecules by colorectal cancer cells can decrease their tumorigenicity greatly, and the B7 molecule can augment the activation of the CTLs against colorectal cancer, and it plays an important role in CTL effector function as well.
文摘背景与目的:EB病毒(Epstein-Barr virus,EBV)感染在鼻咽癌发生中发挥重要作用。本研究的目的在于探讨潜伏的EBV活化后形成的复发感染对鼻咽癌发生过程中全基因组表达的影响。方法:收集EBV阳性(EBV+)/EBV阴性(EBV-)的鼻咽癌靶标基因表达谱和EBV复发感染次数不同的鼻咽癌细胞株基因表达谱,通过生物信息学手段对其进行交叉比较,统计学分析筛选出25个差异基因。运用DAVID(database for annotation,visualization and integrated discovery)、pSTIING(protein,signaling,transcriptional interactions and inflammation networks gateway)、GATHER(gene annotation tool to help explain relationships)和TELiS(transcriptionelement listening system)在线分析工具对25个共同差异基因进行表达模式的预测。结果:与EBV初次感染时的基因表达谱比较,仅有DUSP1、TOP1、HOXA9、DEK、IMPDH2、PABPC1等25个基因在EBV复发感染时仍然呈明显差异表达;这25个差异基因及其相关转录因子主要通过2种模式相互作用:一条主要涉及TOP1、DUSP1、DUSP6和RPS28等,另一条是由PITX1、CD9、HOXA9和IMPDH2组成的转录相关环。结论:EBV潜伏后的复发感染,可能通过筛选到的差异基因的2种表达模式发挥功能,最终导致鼻咽癌的发生。
