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Inhibition of pGPU6/GFP/Neo-hTERT-shRNA on colorectal cancer SW480 cell xenograft in nude mice
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Abstract:
Objective : To investigate the treatment effect of recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA targeting hTERT gene on human colorectal cancer SW480 cell xenograft in nude mice. Methods: Human colorectal cancer SW480 cells were subcutaneously implanted under the skin of the right armpit to establish nude mice model of colorectal cancer, after the tumors grew to a definite size. The mice were randomly divided into three groups: normal saline (NS group), pGPU6/GFP/Neo-NC-shRNA group（NC-shRNA group）and pGPU6/GFP/Neo-hTERT-shRNA group（hTERT-shRNA group）. After each group was treated for 6 consecutive times, the growth status of the tumor was observed, tumor volume was measured, tumor growth curve was drawn，tumor tissue morphology was observed with H-E staining, the expression of hTERT protein in the tumors was detected by immunohistochemistry, cell apoptosis was inspected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL), and the expression of hTERT mRNA was checked by RT-PCR. Results: The growth of tumor volume became slower in hTERT-shRNA group than did that in NS group and NC-shRNA group. Compared with NS group and NC-shRNA group, the tumor cell morphology changed obviously and the number of apoptotic cells increased significantly in the transplanted tumor tissues in hTERT-shRNA group（\[363±505\]% vs \[5.25± 1.06\]%, \[6.95±1.07\]%，P<0.01). Compared with NS group and NC-shRNA group, the expression of hTERT protein was significantly inhibited in hTERT-shRNA group（\[171.42±30.94\] vs \[14689±21.43\], \[13735±25.49\], P<0.01）. Compared with NS group and NC-shRNA group, the expression of hTERT mRNA was significantly inhibited in hTERT-shRNA group （0.39±0.09 vs 0.81±0.34, 0.75±0.21, P<0.05）. Conclusion: Recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA promotes apoptosis of implanted human colorectal cancer by down-regulating the expression of hTERT mRNA and hTERT protein in tumor tissues, thus inhibiting the growth.
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Project Supported:
Project supported by the Natural Science Foundation of Guangxi Zhuang Autonomous Region（No.2010GXNSFA013238）, and the Key Science Foundation of Health Bureau of Guangxi Zhuang Autonomous Region（No.Z200971）
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