论文部分内容阅读
目的外源性人ＴＩＭＰ－２基因在人ＬＡＫ细胞中的表达及其对胃癌浸润转移的抑制作用。方法通过磷酸钙－ＤＮＡ共沉淀的方法将ＴＩＭＰ－２重组真核表达质粒ｐＬ（ＴＩＭＰ－２）ＳＮ转染兼性包装细胞ＰＡ３１７包装形成病毒颗粒。用病毒液感染人ＬＡＫ细胞，再以Ｎｏｒｔｈｅｒｎ杂交、ＳＤＳ－ＰＡＧＥ及反向酶谱试验进行表达及活性分析。用转基因ＬＡＫ细胞（ＬＡＫ／ＬＴ）２×１０６给胃癌转移模型腹腔内注射。结果ＴＩＭＰ－２在ＬＡＫ／ＬＴ中ｍＲＮＡ高度表达，在ＬＡＫ中弱表达。ＬＡＫ／ＬＴ细胞中有相对分子质量２１０００大小的蛋白质表达，具抑制Ⅳ型胶原酶降解明胶的活性。肿瘤种植后１０周，对照组肿瘤体积为（４．００±１．７０）ｃｍ３，肝转移率和腹膜转移率均达１００％，脾脏转移率为７０％（ｎ＝１０）；单用ＬＡＫ细胞治疗组肿瘤体积为（２．７２±１．２０）ｃｍ３，肝及腹膜转移率分别为６０％和７０％；ＴＩＭＰ－２高表达的ＬＡＫ／ＬＴ细胞治疗组肿瘤体积仅（１．２１±１．１６）ｃｍ３，肝转移率与腹膜转移率仅分别为２０％和３０％。肿瘤生长抑制率达７０％，肝及腹膜转移抑制率分别为８０％和７０％。结论ＴＩＭＰ－２对胃癌生长及转移具有抑制作用。
Objective To investigate the expression of exogenous human TIMP-2 gene in human LAK cells and its inhibitory effect on the infiltration and metastasis of gastric cancer. METHODS: The recombinant eukaryotic expression plasmid pL (TIMP-2) SN TIMP-2 was transfected into the packaging cell PA317 by calcium phosphate-DNA co-precipitation to form virus particles. Human LAK cells were infected with virus solution, and then Northern blot, SDS-PAGE and reverse zymography test were performed for expression and activity analysis. The transgene LAK cells (LAK/LT) 2×106 were injected intraperitoneally into gastric cancer metastasis models. Results TIMP-2 was highly expressed in LAK/LT and weakly expressed in LAK. LAK/LT cells express proteins with a relative molecular mass of 21,000 and inhibit the activity of collagen type IV to degrade gelatin. Ten weeks after tumor implantation, the tumor volume in the control group was (4.00±1.70)cm3. The liver metastasis rate and peritoneal metastasis rate were all 100%. The spleen metastasis rate was 70% (n=10); LAK cells were used alone. The tumor volume in the treatment group was (2.72±1.20)cm3, liver and peritoneal metastasis rates were 60% and 70%, respectively; the tumor volume of the TIMP-2 highly-expressing LAK/LT cell treated group was only (1.21±1). .16) In cm3, the liver metastasis rate and peritoneal metastasis rate were only 20% and 30%, respectively. The tumor growth inhibition rate was 70%, and the liver and peritoneal metastasis inhibition rates were 80% and 70%, respectively. Conclusion TIMP-2 can inhibit the growth and metastasis of gastric cancer.