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Recombinant human adenovirus P53 enhances radiosensitivity of human hepatocellular carcinoma in vitro
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Abstract:
Abstract Objective:To investigate the inhibitory effect of recombinant human adenovirus P53(rAdP53) against human hepatocellular carcinoma cell lines with different P53 statuses and its enhancing effect on radiosensitivity. Methods: 〖WTBZ〗Two human hepatocellular carcinoma cell lines with different P53 genetic statuses, PLC/PRF/5 (MT P53) and SMMC7721 (WT P53) were infected with recombinant adenovirus carrying wildtype P53 gene (rAdP53) with increasing multiplicities of infection (MOI). P53 protein expression was detected by Western blotting assay; cell survival was evaluated by MTT; radiosensitivity was evaluated using a clonogenic assay; and cell apoptosis was assayed by TUNEL. Results: Infection efficiency of 50 MOI values of rAdP53 to PLC/PRF/5 and SMMC7721 cells were 89.37% and 86.53%, respectively. The expression of P53 protein was significantly increased in both cell lines 48 h after infection, and cell survival rates of PLC/PRF/5 and SMMC7721 cells were 3.41% and 35.44%, respectively. Inhibitory rates of PLC/PRF/5 and SMMC7721 cells 5 d after infection were 41.91% and 17.03%, respectively. Cells were treated with rAdP53 at 20 MOI for 48 h and then cells were irradiated (4 Gy); the apoptotic rates in PLC/PRF/5 and SMMC7721 cells were (269±5.6)% and (16.4±2.9), respectively (P<0.01). Clone formation assay showed that rAdP53 enhanced sensitivity of PLC/PRF/5 and SMMC7721 cells to radiotherapy, and SER (sensitive enhancement ratio) of Dq were 1.30 and 1.16, respectively, and SER of D0 were 1.57 and 1.25, respectively. Conclusion: rAdP53 can greatly inhibit proliferation, induce l apoptosis and increase radiosensitivity of human hepatocellular carcinoma cells. But the aforesaid efficacy of rAdP53 in PLC/PRF/5 cells is stronger that in SMMC7721 cells, which suggests that hepatocellular carcinoma cells with different P53 statuses responds differently to treatment with rAd-P53 or combination of rAdP53 and irradiation.
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Project Supported:
Supported by the National Natural Sience Foundation of China (No.30872975)
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