Q61R和V112A突变的HRAS基因在NIH小鼠体内诱发肿瘤的实验研究

doi:10.3969/j.issn.1004-616x.2019.01.001

Abstract

Abstract: OBJECTIVE:To evaluate oncogenicity of HRAS (V112A) and its G12C, G12C,Q61R,G12C/G12/Q61R mutants in NIH mice. METHODS:HRAS (V112A) containing the V112A mutation was amplified from the DiFi human colon cancer cells. G12C[HRAS(V112A/G12C)],G12C[HRAS(V112A/G12C)],Q61R[HRAS(V112A/Q61R)] and 3 sites joint[HRAS(V112A/G12C/G12C/Q61R)] mutants were generated with site directed mutation PCR. After HRAS (V112A) and the above 4 mutants were inserted into pCDNA3.1(+) and the in vitro expression was verified with Western blot,each plasmind was injected into 8 6-8 weeks aged female NIH mice intracutaneously with a dosage of 100 μg per mouse to observe the expression of tumorgenicity. Pathological tissues was assayed with PCR and Western blot was used to detect the presence of the HRAS gene and protein. RESULTS:HRAS(V112A) and the 4 designed mutants were amplified successfully. After transfection of L929,HRAS expression was detected. Four months after the injection,4 mice from the HRAS (V112A) injected group developed hyperplastic lesions and 1 mouse from the HRAS(V112A/Q61R) group developed tumor. No lesions were observed in the HRAS (V112A/G12C),HRAS(V112A/G12C) and HRAS(V112A/G12C/G12C/Q61R) groups. PCR and Western blot results show the existence and expression of the corresponding HRAS genes. CONCLUSION:HRAS containing the V112A mutation could induce hyperplastic lesions in mice,while HRAS containing V112A and Q61R mutations could induce tumors in NIH mice. The HRAS(V112A) containing G12C,G12C and G12C/G12C/Q61R did not induce observable lesions.

Key words: HRAS, point mutation, oncogenicity, mouse tumor formation experiment

CLC Number:

R73-35+4

ZHANG Feng, ZHAO Long, FAN Jinping, WU Xueling, MENG Shufang. Induction of tumor in NIH mice by the plasmid containing the V112A and Q61R mutated HRAS[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 1-8.

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Induction of tumor in NIH mice by the plasmid containing the V112A and Q61R mutated HRAS

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