IL-15基因转染人CIK细胞过继转移治疗胃癌的实验研究
发布时间：2018-11-24 11:30
【摘要】： 本实验构建了hIL-15-IRES-TK逆转录病毒载体,并证实可有效转染CIK细胞。其转染可使CIK细胞自分泌白介素-15(IL-15)。鉴于逆转录病毒转染具有潜在的导致自身免疫疾病的危险,同时表达的胸苷激酶基因(HSV-TK)可使被转染的CIK细胞在丙氧乌苷(GCV)的处理下能被有效清除。这为hIL-15-TK病毒载体应用于临床CIK细胞过继转移治疗肿瘤的安全性提供了重要的基础。 为了观察hIL15-IRES-TK质粒转染的人源T细胞在体内治疗人源胃肠道肿瘤的疗效,我们利用T细胞免疫缺陷小鼠(Balb/c-nu/nu鼠)以人的低分化胃腺癌细胞株BGC-823成功制备了人源胃癌细胞荷瘤小鼠模型,分别以hIL-15-IRES-TK逆转录病毒转染和未转染的CIK细胞在瘤体局部注射,1次/周,共2周,治疗后观察2周。以生理盐水局部注射作为空白对照组。每周测量裸鼠肿瘤体积1次,4周后处死裸鼠,取出肿瘤组织称重。 经过2周的治疗,分别注射hIL-15-IRES-TK质粒转染与非转染的CIK细胞的两组荷瘤小鼠的肿瘤体积出现了显著性差异,显示出hIL-15-IRES-TK质粒转染的CIK细胞比非转染的CIK细胞有更强的抑制肿瘤增长的活性。在第3、4周,停止CIK治疗后3组小鼠的肿瘤体积均有所增加,但hIL-15-IRES-TK质粒转染与非转染的CIK细胞组均小于生理盐水组,说明CIK细胞过继转移治疗具备一定的抑制肿瘤细胞的作用。而hIL-15-IRES-TK质粒转染与非转染的CIK细胞两组小鼠的肿瘤体积差异依然显著,显示出hIL-15-IRES-TK质粒转染的CIK细胞比非转染的CIK细胞抑制肿瘤增长的时效更长。 在实验过程中IL-15转染CIK组开始治疗后有两只裸鼠肿瘤溃烂,第二周开始溃烂结疤,第四周溃烂愈合,肿瘤消失。对照组与CIK组则无此现象出现。这一现象也侧面证实了IL-15-IRES-TK质粒转染的CIK细胞比非转染的CIK细胞具备更强和更持续的杀瘤效应。 结论:实验表明CIK细胞可以有效抑制胃腺癌细胞株BGC-823在体内的生长,而hIL-15-IRES-TK质粒转染的CIK细胞不仅可以抑瘤,还可以致肿瘤消退。
[Abstract]:In this study, hIL-15-IRES-TK retrovirus vector was constructed, and it was confirmed that it could be transfected into CIK cells effectively. The transfection can make CIK cells secrete interleukin-15 (IL-15). In view of the potential risk of autoimmune disease caused by retrovirus transfection, the expressed thymidine kinase gene (HSV-TK) can effectively remove the transfected CIK cells under the treatment of (GCV). This provides an important basis for the safety of hIL-15-TK virus vector in clinical CIK cell adoptive metastasis. To observe the therapeutic effect of human T cells transfected with hIL15-IRES-TK plasmid on human gastrointestinal carcinoma in vivo. We successfully established a tumor-bearing mouse model of human gastric cancer cells by using T cell immunodeficient mice (Balb/c-nu/nu mice) with human poorly differentiated gastric adenocarcinoma cell line BGC-823. HIL-15-IRES-TK retrovirus-transfected and untransfected CIK cells were injected locally into the tumor once a week for 2 weeks. Saline injection was used as the blank control group. The tumor volume of nude mice was measured once a week. After 4 weeks, the nude mice were killed and the tumor tissue was weighed. After two weeks of treatment, there was a significant difference in tumor volume between the two groups injected with hIL-15-IRES-TK plasmid and non-transfected CIK cells, respectively. The results showed that CIK cells transfected with hIL-15-IRES-TK plasmid had stronger inhibitory activity against tumor growth than non-transfected CIK cells. At the 3rd week, the tumor volume of the three groups increased after CIK treatment, but the hIL-15-IRES-TK plasmid transfection and non-transfection CIK cells were smaller than the normal saline group. These results suggest that adoptive transfer therapy of CIK cells has a certain inhibitory effect on tumor cells. However, the difference of tumor volume between hIL-15-IRES-TK plasmid transfected and non-transfected CIK cells was still significant, indicating that hIL-15-IRES-TK plasmid transfected CIK cells had a longer time to inhibit tumor growth than non-transfected CIK cells. During the course of the experiment, two nude mice had tumor ulceration after the IL-15 transfection CIK group began treatment, the second week began to ulcerate scarring, the fourth week ulcerated and healed, the tumor disappeared. There was no such phenomenon in the control group and CIK group. This phenomenon also confirmed that CIK cells transfected with IL-15-IRES-TK plasmid had stronger and more sustained tumor killing effect than non-transfected CIK cells. Conclusion: CIK cells can effectively inhibit the growth of gastric adenocarcinoma cell line BGC-823 in vivo, and hIL-15-IRES-TK plasmid transfected CIK cells can not only inhibit tumor, but also cause tumor regression.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
本文编号：2353540
[Abstract]:In this study, hIL-15-IRES-TK retrovirus vector was constructed, and it was confirmed that it could be transfected into CIK cells effectively. The transfection can make CIK cells secrete interleukin-15 (IL-15). In view of the potential risk of autoimmune disease caused by retrovirus transfection, the expressed thymidine kinase gene (HSV-TK) can effectively remove the transfected CIK cells under the treatment of (GCV). This provides an important basis for the safety of hIL-15-TK virus vector in clinical CIK cell adoptive metastasis. To observe the therapeutic effect of human T cells transfected with hIL15-IRES-TK plasmid on human gastrointestinal carcinoma in vivo. We successfully established a tumor-bearing mouse model of human gastric cancer cells by using T cell immunodeficient mice (Balb/c-nu/nu mice) with human poorly differentiated gastric adenocarcinoma cell line BGC-823. HIL-15-IRES-TK retrovirus-transfected and untransfected CIK cells were injected locally into the tumor once a week for 2 weeks. Saline injection was used as the blank control group. The tumor volume of nude mice was measured once a week. After 4 weeks, the nude mice were killed and the tumor tissue was weighed. After two weeks of treatment, there was a significant difference in tumor volume between the two groups injected with hIL-15-IRES-TK plasmid and non-transfected CIK cells, respectively. The results showed that CIK cells transfected with hIL-15-IRES-TK plasmid had stronger inhibitory activity against tumor growth than non-transfected CIK cells. At the 3rd week, the tumor volume of the three groups increased after CIK treatment, but the hIL-15-IRES-TK plasmid transfection and non-transfection CIK cells were smaller than the normal saline group. These results suggest that adoptive transfer therapy of CIK cells has a certain inhibitory effect on tumor cells. However, the difference of tumor volume between hIL-15-IRES-TK plasmid transfected and non-transfected CIK cells was still significant, indicating that hIL-15-IRES-TK plasmid transfected CIK cells had a longer time to inhibit tumor growth than non-transfected CIK cells. During the course of the experiment, two nude mice had tumor ulceration after the IL-15 transfection CIK group began treatment, the second week began to ulcerate scarring, the fourth week ulcerated and healed, the tumor disappeared. There was no such phenomenon in the control group and CIK group. This phenomenon also confirmed that CIK cells transfected with IL-15-IRES-TK plasmid had stronger and more sustained tumor killing effect than non-transfected CIK cells. Conclusion: CIK cells can effectively inhibit the growth of gastric adenocarcinoma cell line BGC-823 in vivo, and hIL-15-IRES-TK plasmid transfected CIK cells can not only inhibit tumor, but also cause tumor regression.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
【引证文献】
相关硕士学位论文 前1条
1 甘正艺;乌司他丁对大鼠光气所致急性肺损伤的抑制及对IL-15和ICAM-1的影响[D];复旦大学;2012年
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