cbl-b基因转导淋巴细胞对人前列腺癌细胞LNCaP杀伤作用的实验研究

Experimental study on cytotoxicity of lymphocytes transfected with cbl-b gene in prostate carcinoma cells LNCaP

目的观察将cbl-b基因导入淋巴细胞能否增强其对人前列腺癌细胞LNCaP的杀伤作用。方法 cbl-b基因淋巴细胞组采用脂质体法将cbl-b基因导入淋巴细胞,在蛋白印记法证实导入基因成功表达后,再用CCK-8法检测其对前列腺癌的杀伤作用,并与正常人外周血中分离淋巴细胞组比较。结果 12h、24h、48h三个时间段,转基因淋巴细胞的杀伤率在10:1、20:1、40:1三种浓度与单纯淋巴细胞比较,杀伤率明显高于单纯淋巴细胞组,差异均有统计学意义(F分别=74.25、1745.90、1637.45,P均<0.05)。其中转cbl-b基因淋巴细胞组在10:1和40:1浓度时24h杀伤率明显高于12h、48h,差异均有统计学意义(t分别=18.37、36.00、17.25、35.21,P均<0.05),但12h、48h的杀伤率比较,差异无统计学意义(t分别=1.26、2.51,P均>0.05)。在20:1时12h杀伤率明显低于24h、48h,差异均有统计学意义(t分别=16.25、19.12,P均<0.05),但24h、48h的杀伤率比较,差异无统计学意义(t=1.06,P>0.05)。且在40:1转染24h杀伤作用最强,可达(44.16±0.67)%。结论转导入cbl-b基因可增强淋巴细胞对人前列腺癌细胞LNCaP的杀伤作用,且杀伤率最强的比例与时间段为40:1、24h。

Objective To investigate whether lymphocytes transfected with cbl-b gene are capable of improving the cytotoxicity against prostate carcinoma cells LNCaP in vitro.Methods Lymphocytes cells were isolated from normal human peripheral blood and were cultured in vitro.The lymphocytes were transfected with cbl-b gene by liposome mediated gene transfection.The protein expression was determined with Western blot.The cytotoxicity of lymphocytes against prostate cancer cells were examined by CCK-8 assay,and compared with peripheral blood lymphocytes isolated group.Results Compared with lymphocytes cell group,every cell group transfected significantly enhanced the killing effect at 12 hours,24 hours and 48 hours.There was a significant difference in the different concentration.The group 10：1（F=74.25,P0.05）,the group 20：1（F=1745.90,P0.05）,the group 40：1（F=1637.45,P0.05）.Further comparing the results according to the time,there was statistically significant difference between 24 hours and 12 hours,24 hours and 48 hours in the concentration of 10：1 and 40：1（t=18.37,36.00,17.25,35.21,P0.05）,but there was no significant difference,comparing with 12 hours and 48 hours（t=1.26,2.51,P0.05）.There was statistically significant difference between 12 hours and 24 hours,12 hours and 48 hours in the concentration of 20：1（t=16.25,19.12,P0.05）,but there was no statistically significance at 24 hours and 48 hours（t=1.06,P0.05）.In the condition of 40：1 transfected 24 hours,the killing rate was up to（44.16±0.67）%.Conclusions It is possible to improve the cytotoxicity of lymphocytes against prostate carcinoma cells LNCaP in vitro by being transfected with cbl-b gene,especially for the concentration 40：1 and the time period of 24 hours.