摘要
本室曾报道ＲＡ５３８转染亲本食管癌细胞系ＥＣ８７１２，可使后者发生终末分化和凋亡。为进一步研究这个ｃＤＮＡ的生物学作用，用ＲＡ５３８０．３ｋｂ的片段与ｎｅｏ基因构建了一个ｐＲＡ５３８－０．３－ｎｅｏ表达质粒，并转入三种不同的人癌细胞系即ＥＣ８７１２、ＨＬ６０和ＧＬＣ（一个来自人肺腺癌的细胞系）。经含Ｇ４１８培养液筛选后，进行了生长速度、３Ｈ－ＴｄＲ掺入率、细胞形态学、在半固体琼脂中克隆形成能力，以及裸鼠异种接种等检测。对获得的抗Ｇ４１８细胞群体均用上述０．３ｋｂ片段进行细胞原位杂交以证明其掺入和表达。同三个亲本癌细胞系比较这些细胞群体的生长和恶性表型均受到明显的抑制。
As previously reported, transfer of RA538 into parental esophageal cancer EC8712 cell line induced its terminal differentiation and apoptosis. To further study the biological effects of this cDNA,an expression plasmid containing an insert of the putative coding fragment(about 0.3kb)of RA538 and neo resistance gene was constructed(designated pRA538-0.3 neo)and transferred into three different human cancer cell lines:EC8712,HL60 and GLC,cell line derived from an adenocarcinoma of the lung. After selection in G418-containing cul ture media,the growth rate, 3H-thymidine incorporation rate,cell morphology,olony-form ation in soft-agar, and heterotransplantation into nude mice of the surviving cell populations were tested. In situ hybridization verified the uptake and expression of the 0.3kb fragment of RA538 in these G418 resistant cell populations.Significant reduction in growth rate and sup pression of malignant phenotype were observed in all these cells in comparison with their parental cancer cell lines.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1994年第4期245-250,共6页
Acta Academiae Medicinae Sinicae
基金
"863"高技术
医科院重点项目
博士后基金
