摘要
过去有人提出EBV—DNA是通过病毒颗粒感染,或者是携带EBV基因的细胞与宿主细胞融合这两种途径之一,导致病毒基因转输入宿主细胞DNA。近年我们注意到:基因片段转染,是基因转移和导致细胞转化的一种重要形式。 本实验应用DNA介导的基因转移的钙沉淀改良法,在体外培养将EBV重复序列区的BamHl—W及其相邻片段转染宿主细胞(大鼠成纤维细胞,Rat—1),3周内出现转化灶,经半固体培养基进行转化选择,从而获得多株高效转化克隆,其中W_4和W_(10)通过裸鼠体内接种与鼠间移植,最后建立高移植成功率(94—100％)的可移植性裸鼠瘤(CH—1,CH—2)。 经SP6/T7RNA多聚酶诱导RNA探针、缺口翻译DNA探针和亚细胞定位的原位杂交探针综合应用,测知经EBV基因片转转染后多次传代的转化细胞(W_(10),W_4)和多次移植的裸鼠瘤(CH—1,CH—2)DNA中存在着EBV基因片段。另一方面,测知人类的鼻咽癌组织和Burkitt淋巴瘤细胞株以及绒猴的EBV转化细胞株中,恒定地整合着该基因片段。因此,EBV基因片段的活性及其在癌变过程中的作用,与癌基因激活等问题值得进一步研究。
Cells of rat fibroblast cell line (Rat-1) Were transfected with EBV-BamH 1W and adjacent fragments using Calcium precipitation method modified by us. Within three weeks after the tranfection typical transformed cell foci have been found on soft agar cultures. Afer selecting cuture we obtained several high efficient transformed clones including WTO and W4 . Using the tumor formation experiment, the transformed clones including WlO and W4 have been implanted subcutaneously to nude mice (N C strain) and grewup transpla-ntabl tumors, xeno-implantation sarcomas CH-1 and CH-2 , in vivo with a high transplantable rate of 94- 100 %.After the comprehensive use of SP6/T7 RNA polymerase inducing Riboprobes, nick translation probes and probes of whole tissue hybridization In Situ for subcellular location , it is proven that EBV-BamHI DNA fragments existed not only in the host cell DNAs of repeti live subculture transformed foci but also in the tumor cell DNAs and chromosomes of transplantable nude mice tumors. On the other hand, the commparative analysis on this Rat-1 tumor induced by the sugbgenomic fragment and human cancer, biopsies of nasopharyngeal carcinoma and cell lines of Burkitt lymphoma, showed that the DNAs extracted form both cancer have steadily integrated with EBV-DNA framgents.The result indicates that the biological activity of EBV gene fragment and its coation with oncogene for the malignant transformation of host cells are worthy of the further study.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
1989年第1期9-12,共4页
Chinese Journal of Cancer
基金
国家自然科学基金
广东省高教局基因工程研究基金
国家科委七
五攻关资助
