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目的ＴＮＦ存在两种形式，目前研究集中于分泌型ＴＮＦ（ｓ－ＴＮＦ），对跨膜型ＴＮＦ（ＴＭ－ＴＮＦ）的研究很少，但仅有的资料表明，ＴＭ－ＴＮＦ较ｓ－ＴＮＦ有更广的杀瘤谱，故本实验拟初步探讨两型ＴＮＦ杀瘤效应的差异。方法应用生物活性测定法检测两型ＴＮＦ的胞毒作用，ＲＴ－ＰＣＲ检测靶细胞ＴＮＦＲｍＲＮＡ表达水平，ＤＮＡ梯带、ＴＵＮＥＬ和Ｓｕｂ－Ｇ１峰检测细胞凋亡，所得结果全部采用ｔ检验。结果ＴＭ－ＴＮＦ不仅能杀伤ｓ－ＴＮＦ敏感细胞株，而且能有效杀伤ｓ－ＴＮＦ耐受细胞（ＨＬ－６０，Ｋ５６２，ＨｅｐＧ２，Ｒａｊｉ）（Ｐ＜０．０１）；ＲＴ－ＰＣＲ检测试验所用的６株靶细胞，其中３株为ＴＮＦＲⅠ单表达（Ｌ９２９，ＭＣＦ－７和ＨｅｐＧ２），２株为ＴＮＦＲ双表达（ＨＬ－６０和Ｋ５６２），１株为ＴＮＦＲⅡ单表达（Ｒａｊｉ）；ｓ－ＴＮＦ敏感细胞单表达ＴＮＦＲⅠ，ＴＭ－ＴＮＦ可杀伤表达任何类型ＴＮＦＲ的细胞；ＴＭ－ＴＮＦ以诱导细胞凋亡为主（Ｐ＞０．０１），ｓ－ＴＮＦ以诱导坏死为主（Ｐ＜０．０１）。结论两型ＴＮＦ的胞毒作用存在显著性差异，由于ＴＭ－ＴＮＦ在局部发挥作用且以诱导凋亡为主，杀瘤谱较ｓ－ＴＮＦ更广，故在肿瘤治疗中可能优于ｓ－Ｔ?
Objective TNF exists in two forms. At present, the research focuses on secretory TNF (s-TNF). There are few studies on transmembrane TNF (TM-TNF), but the only data shows that TM-TNF is more effective than s-TNF. The broader spectrum of killing tumors, so this experiment is intended to tentatively explore the differences between the two types of TNF killing effect. Methods The cytotoxicity of two types of TNF was detected by biological activity assay. The expression of TNFR mRNA in target cells was detected by RT-PCR. Apoptosis was detected by DNA ladder, TUNEL and Sub-G1 peaks. All the results were analyzed by t test. Results TM-TNF could not only kill s-TNF-sensitive cell lines, but also effectively kill s-TNF-resistant cells (HL-60, K562, HepG2, Raji) (P<0.01); RT-PCR assay used Among 6 target cells, 3 were TNFRI single (L929, MCF-7 and HepG2), 2 were TNFR double (HL-60 and K562), 1 was TNFRII single (Raji), and s-TNF was sensitive Cells expressed TNFRI alone, TM-TNF could kill cells that expressed any type of TNFR; TM-TNF induced apoptosis mainly (P>0.01), and s-TNF mainly induced necrosis (P<0.01). . Conclusion There is a significant difference in the cytotoxic effect of the two types of TNF. Because TM-TNF plays a role locally and induces apoptosis, the tumor killing spectrum is more extensive than s-TNF, so it may be superior to s-T in the treatment of tumors. ?