转基因溶瘤性单纯疱疹病毒对人类乳腺癌细胞及其干细胞的溶瘤作用

 【目的】 通过体外实验观察溶瘤性单纯疱疹病毒(HSV)载体G47Δ对人类原代乳腺癌细胞及乳腺癌干细胞毒效应,探讨其对人乳腺癌的治疗作用。【方法】 选取6例浸润性导管癌组织,体外培养人原代乳腺癌细胞及人原代乳腺癌干细胞,将G47Δ按不同感染复数(MOI),即MOI=0.01和MOI=0.1加入各实验组中,观察每天细胞的生长情况。G47Δ含有LacZ基因,其表达产物可被X-gal染色检测。 【结果】 G47Δ病毒在乳腺癌细胞中可复制和传播通过X-gal染色法得以证实。 G47Δ对人原代乳腺癌细胞及乳腺癌干细胞都具有杀伤作用,对乳腺癌细胞更加敏感。在MOI=0.01组及0.1组中,感染病毒后第4 d,人原代乳腺癌细胞被杀灭的比例分别是91%及96%,乳腺癌干细胞被杀灭的比例分别是43%及78%;感染病毒后第6天,两组中人原代乳腺癌细胞接近全部被杀灭,乳腺癌干细胞被杀灭的比例分别是92%、98%。 【结论】 G47Δ可有效杀灭人乳腺癌细胞及其干细胞;乳腺癌细胞比乳腺癌干细胞对G47Δ更具有敏感性。

Effect of Transgenic Oncolytic Herpes Simplex Virus Vectors on Human Breast Cancer and Its Stem Cells

【Objective】To investigate the efficacy of oncolytic herpes simplex virus (HSV) vector, G47Δ for the treatment of the human breast cancer through the observation of cytotoxicity on human primary breast cancer cells and its stem cells in vitro. 【Methods】 Human primary breast cancer cells and breast cancer stem cells from 6 breast invasive ductal carcinoma samples were cultured and inoculated with G47Δ of two different multiplicities of infection (MOI), MOI=0.01 and MOI=0.1. The condition of cell growth was observed every day. G47Δ contained the LacZ gene, and expression in infected cells was detected with X-gal histochemistry. 【Results】 It was confirmed that G47Δ can multiply and disseminate in breast cancer cells with X-gal histochemistry. G47Δ was cytotoxic for human primary breast cancer cells and breast cancer stem cells in vitro, but the cytotoxicity for human primary breast cancer cells was stronger than its stem cells. X-gal staining of infected tumor cells in vitro illustrated the replication and spread of the virus. In the MOI=0.01 group and 0.1 group, on the fourth day after infection, 91% and 96% human primary breast cancer cells were killed, 43% and 78% human primary breast cancer stem cells were killed. In the MOI = 0.01 and MOI = 0.1 group, on the sixth day after infection, the killing rates of human primary breast cancer cells were nearly 100% in both groups; the killing rates of human primary breast cancer stem cells were 92% and 98%, respectively. 【Conclusions】 Oncolytic herpes simplex virus (HSV)vector G47Δ was effective at killing human primary breast cancer cells and breast cancer stem cells in vitro. Breast cancer cells were more sensitive to G47Δ than its stem cells.