摘要
目的探讨PUMA基因转染抑制胰腺肿瘤生长的体内外效果。方法利用脂质体转染法将表达PUMA的质粒转染导入胰腺癌细胞株PC-3中,G418筛选出阳性克隆,Western和RT-PCR法检测PUMA转染后PC-3PUMA的表达,流式细胞仪检测转染后细胞凋亡率;分别将转染PUMA的PC-3细胞(实验组)和未转染的PC-3细胞移植到裸鼠体内,比较裸鼠移植肿瘤的大小和重量以及PUMA表达。结果PUMA表达质粒转染的PC-3细胞(PC-3/PUMA)稳定表达PUMA,其细胞凋亡率为(5.50±0.90)%,明显高于未转染组的(1.073±0.248)%和空载体转染组的(1.08±0.35)%(P<0.05);裸鼠接种4周后PC-3/PUMA细胞成瘤率为70%,PC-3细胞和空载体PC-3细胞成瘤率为100%(P>0.05),PC-3/PUMA细胞形成的肿瘤体积比PC-3细胞和空载体PC-3细胞明显减小(P<0.05),并且形成的肿瘤组织中PUMA高表达。结论胰腺癌细胞中缺失PUMA基因表达,PUMA转染胰腺癌细胞后表达PUMA,并能促进细胞凋亡。
Objective To investigate the antitumor effect of PUMA gene transfection into pancreatic cancer cell line PC-3. Methods Plasmid PUMA-pEGFP C1 was introduced into pancreatic cancer cell line PC-3 by LipofectinamineTm 2000 transfection. Positive clones were screened by G418 and stable expression of PUMA was detected by Western blot and RT PCR method. FCG was performed to test the rate of apoptosis. Fifteen athymic mice were equally randomized into 3 groups： the experiment group xenografted with PUMA expressing cells, the empty vector control group, and the control group. Weight of the mice and size of the tumors were measured every week. Eight weeks later the mice were killed and the tumors were removed. The tumor volume and weighl were compared between the three groups. Western blot method was used to determine the expression of PUMA genes. Results The expression of PUMA in stable transfection PC 3 cells was detected. The rale of apoptosis by FCG was significantly higher in the experimental group （5.50 ± 0. 90%） compared with the empty vector control group （1. 073 ± 0. 248%） and the control group （1. 08 ± 0. 35%）. There was no significanl difference in lumorigenicity between PC-3/PUMA and PC-3 cells （70% vs 100%）, though the tumor volume generated by PC 3/PUMA cells was significantly smaller（P 〈 0.05）. There was no significant difference in all the indicators between the empty vector control group and the control group （both P 〉 0.05）. The expression of PUMA protein in the experimental group was higher than that in the empty vector control group and control group. Conclusions Rexpression of PUMA gene, the expression of which is lost in human pancreatic adenocarcinoma, induces apoptosis, thus inhibiling tumor growth.
出处
《胰腺病学》
2006年第6期355-357,共3页
Chinese JOurnal of Pancreatology
关键词
胰腺肿瘤
PUMA基因
转染
细胞凋亡
Pancreatic neoplasms PUMA（p53 up regulaled modulator of apoptosis） gene
Transfeclion
Apoptosis
