CAJ | 학술논문

目的：构建癌基因H-ras真核表达载体，并检测其对肿瘤细胞生长的作用。方法：以乳腺文库为模板，PCR扩增H-ras基因片段，将其插入pXJ-40-myc载体后转染人胚肾HEK293T细胞，用Western印迹检测该融合蛋白的表达；将重组质粒转染人肠癌HCT-116细胞和人肝癌HepG2细胞，通过细胞生长曲线（CCK8法）对myc-H-ras影响肿瘤细胞生长的情况进行分析。结果：利用PCR技术，从乳腺文库中扩增出H-ras基因片段，并成功插入pXJ-40-myc载体；重组质粒转染HEK293T细胞，经Western印迹检测融合蛋白正确表达；细胞生长曲线结果显示，转染myc-H-ras的结肠癌HCT-116细胞和人肝癌HepG2细胞比转染空载体的细胞生长快。结论：构建了人H-ras基因真核表达载体，为进一步研究ras基因在肿瘤细胞中的作用奠定了一定基础。
목적：구건암기인H-ras진핵표체재체，병검측기대종류세포생장적작용。방법：이유선문고위모판，PCR확증H-ras기인편단，장기삽입pXJ-40-myc재체후전염인배신HEK293T세포，용Western인적검측해융합단백적표체；장중조질립전염인장암HCT-116세포화인간암HepG2세포，통과세포생장곡선（CCK8법）대myc-H-ras영향종류세포생장적정황진행분석。결과：이용PCR기술，종유선문고중확증출H-ras기인편단，병성공삽입pXJ-40-myc재체；중조질립전염HEK293T세포，경Western인적검측융합단백정학표체；세포생장곡선결과현시，전염myc-H-ras적결장암HCT-116세포화인간암HepG2세포비전염공재체적세포생장쾌。결론：구건료인H-ras기인진핵표체재체，위진일보연구ras기인재종류세포중적작용전정료일정기출。
Objective: To construct the eukaryotic expression vector of myc-tagged human H-ras and detect its function in cancer cells. Methods: Human H-ras gene was amplified from the mammary library by the technique of PCR, and was inserted into eukaryotic expression vector of pXJ-40-myc. The recombinant plasmid pXJ-40-myc-H-ras was identified by sequencing, and transfected into HEK293T cells. The myc-H-ras protein was detect?ed by Western blot. Then CCK8 assay was performed to investigate the effect of myc-H-ras on proliferation of co?lon cancer cell HCT-116 and liver cancer cell HepG2. Results: The DNA fragment of about 600 bp was success?fully amplified by PCR, cloned into pXJ-40-myc, and identified by sequencing. Western blot showed the expres?sion of myc-H-ras in the HEK293T cells. Cell growth curve demonstrated that cells transfected with myc-H-ras grew significantly faster than those transfected with empty vector. Conclusion: The eukaryotic expression vector of myc-H-ras was successfully constructed, which laid a foundation for the study of H-ras in cancer development and progression.