乏氧射线双调控的TK腺病毒载体对乳腺癌细胞生长和凋亡的影响

Influence of TK adenovirus vector regulated by hypoxia and radiation on the growth and apoptosis of breast cancer cells

目的:观察乏氧射线双调控的TK腺病毒载体-Ad.HRE.CArG.HSV-TK对体外培养的乳腺癌细胞Bcap37和MDA-MB-435生长和凋亡的影响。方法:MTT法检测腺病毒转导联合放疗对常氧(37℃,19%O2,5%CO2)和乏氧(37℃,0.1%O2,5%CO2)状态下细胞的生长抑制作用,Annexin V-FITC染色检测细胞凋亡。结果:HSV-TK/GCV系统对乳腺癌细胞具杀伤效应和旁观者效应,Bcap37和MDA-MB-435细胞对GCV的IC50分别为9.09和7.83μg/mL。常氧状态下腺病毒载体(Ad)与放疗(RT)联用组(Ad+RT)细胞生长抑制率和凋亡率明显高于Ad组和相应同等剂量放疗组(P<0.05),对Bcap37和MDA-MB-435细胞的增敏比(SER)均为1.55。乏氧和(或)放疗处理后,乏氧+Ad+RT组抑制率明显高于常氧+Ad+假照射组和常氧+Ad+RT组,P<0.05。结论:Ad.HRE.CArG.HSV-TK腺病毒载体联合放疗可显著抑制乳腺癌细胞生长并诱导凋亡,为进一步开展乳腺癌的基因-放射治疗研究奠定了基础。

OBJECTIVE： To observe the influence of TK ad enovirus vector-Ad. HRE. CArG. HSV TK regulated by hypoxia and radiation on the growth and apoptosis of breast cancer cells in vitro. METHODS： MTT assay was used to detect the growth inhibitory effect of adenovirus transduction on cells under normoxia （37℃,19% O2, 5% CO2） and hypoxia （37 ℃, 0. 1% 02, 5% CO2 ）. Apoptosis was detected by Annexin V-FITC staining. RESULTS： The system of HSV TK/GCV have killing effect and by stander effect on breast cancer cells. IC50 of GCV were 9.09 and 7.83 μg/mL in Bcap37 and MDA MB-435 cells, respectively. Under normoxia,the rate of growth inhibition and apoptosis in group of adenovirus vector（Ad）combined with radiotherapy （RT） was significantly higher than that in Ad group and the same dose of RT group, P〈0.05. The ratios of sensitizer en hencement （SER） were both 1.55. After hypoxia and/or radiation treatment, the inhibition rate in group of hypoxia＋Ad＋RT were obviously higher than that in group of normoxia＋Ad＋fake irradiation and normoxia ＋Ad＋ RT, P〈0.05. CONCLUSIONS： Adenovirus vector of Ad. HRE. CArG. HSV TK combined with radiotherapy can significant suppress the growth and induce apop tosis in breast cancer cells. It provids foundation for further gene radiation therapy of breast cancer.