Effect of oncolytic virus loaded with MAGE-A10 / hβD-2 double gene on bladder cancer cell proliferation and tumor growth#br#
ZHANG Jianjun1 CAI Longjun1 CAI Weiqi2
1.Kewen College, Jiangsu Normal University, Jiangsu Province, Xuzhou 221132, China;
2.Department of Urology, the Affiliated Suqian Hospital of Xuzhou Medical University, Suqian Hospital of Nanjing Drum-Tower Hospital Group, Jiangsu Province, Suqian 223800, China
Abstract：Objective To explore the effect of oncolytic virus SG502-MAGE-A10-hβD-2 loaded with melanoma-associated antigen A10 (MAGE-A10) and human beta defensin 2 (hβD-2) genes on the proliferation and tumor growth of bladder cancer cell lines. Methods The oncolytic virus SG502-MAGE-A10-hβD-2 loaded with MAGE-A10 and hβD-2 genes was constructed. Virus titer was determined by 50% tissue culture infectious dose. CCK-8 method were used to determine the effects of oncolytic viruses SG502 and SG502-MAGE-A10-hβD-2 on the proliferation of T24 and MB49 cells. SG502 group was added with SG502 virus, SG502-MAGE-A10-hβD-2 group was added with SG502-MAGE-A10-hβD-2 virus, and the control group was added with PBS for 24, 48, 72, 96 h, respectively. The MB49 cell was used to construct a C57BL/6 mouse subcutaneously transplanted tumor model. After the tumor grew to 100-150 mm3, the eighteen mice were divided into control group, SG502 group and SG502-MAGE-A10-H βD-2 group according to the random number table method, with six mice in each group. The control group was given 50 μl normal saline for intratumoral multipoint injection. SG502 group was given 50 μl normal saline solution containing SG502 virus with 2×108 virus particles for intratumoral multipoint injection. SG502-MAGE-A10-hβD-2 group was given 50 μl normal saline solution containing SG502-MAGE-A10-hβD-2 virus particles of 2×108 in intratumoral multipoint injection, two times a week. The tumor volume, the expression of MAGE-A10 and hβD-2 proteins in tumor-forming tissues, the expression of Ki67 and the contents of TNF-α and IFN-γ in serum were detected five weeks after intratumoral injection. Results The oncolytic virus SG502-MAGE-A10-hβD-2 was successfully constructed, and the titer of the purified virus was 2.05×1010 PFU/ml. The inhibition rates of T24 and MB49 cells at each time point in SG502 group and SG502-MAGE-A10-hβD-2 group were statistically significant (P ＜ 0.05). There were statistically significant differences in T24 and MB49 cell inhibition rates between SG502 group and control group at 24, 48, 72, and 96 h (P ＜ 0.05). The inhibition rates of T24 and MB49 cells in SG502-MAGE-A10-hβD-2 group and SG502 group at 24, 48, 72, and 96 h were significantly different (P ＜ 0.05). The tumor volume of SG502 group and SG502-MAGE-A10-hβD-2 group at each time point was significantly different (P ＜ 0.05). There were statistically significant differences in tumor volume between SG502 group and control group at 14, 21, 28, 35, and 42 days (P ＜ 0.05). There were statistically significant differences in tumor volume between SG502-MAGE-A10-hβD-2 group and SG502 group at 14, 21, 28, 35, and 42 days (P ＜ 0.05). The tumor weight of SG502 group was lower than that of the control group, and that of SG502-MAGE-A10-hβD-2 group was lower than that of SG502 group, the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01). The levels of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in SG502 group were higher than those in control group, and the levels of TNF-α and IFN-γ in SG502-MAGE-A10-hβD-2 group were higher than those in SG502 group, the differences were highly statistically significant (P ＜ 0.01). After five weeks of intervention, MAGE-A10 and hβD-2 proteins were expressed in SG502-MAGE-A10-hβD-2 group, but not in the control group and SG502 group. The expressions of Ki67 in tumor tissues of SG502 group and SG502-MAGE-A10-hβD-2 group were lower than that of control group, and the differences were highly statistically significant (P ＜ 0.01). Conclusion SG502-MAGE-A10-hβD-2 can inhibit the proliferation of T24 and MB49 cells, inhibit the tumor growth of MB49 tumor-forming mice, and stimulate the secretion of TNF-α and IFN-γ in mice.
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