摘要
目的:建立稳定表达反义表皮生长因子受体(EGFR)的人结肠癌细胞系,研究细胞的生物学行为.方法:应用脂质体介导法将重组克隆携带反义EGFR的逆转录病毒载体转染人结肠癌细胞系LST174,利用细胞计数、生长曲线、MTT法及软琼脂集落形成率测定细胞生长、增殖及恶性表型的转化.结果:转染稳定后的细胞易脱落,细胞膜毛糙,细胞生长呈小片状,不密集.细胞继续培养后,LST174/AS-EGFR组细胞生长受到抑制,生长曲线较LST174组速度减慢,差别显著(A值:0.322±0.014vs0.422±0.033,P<0.05);细胞生长抑制率为23.7%,LST174/AS-EGFR组细胞几乎失去了集落形成能力,且少、松散,而对照组呈叠落堆积生长.结论:反义EGFR对内源性EGFR自分泌因子表达的阻断可抑制结肠癌细胞的生长、增殖及恶性表型的转化.
AIM： To construct a stable cell line through transfecting reverse transcription virus carrier with antisense epidermal growth factor receptor （AS-EGFR） into human colonic cancer cell line （LST174） and investigate its biological behavior.
METHODS： Reverse transcription virus carrier with AS-EGFR was transfected into human colonic cancer cell line LST174 by lipid mediating method. The cellular growth and transformation of malignant phenotype were detected by cellular counting, growth curve, MTT assay and rate of clone formation on soft agar.
RESULTS： The cells after stable transfection exfoliated easily with rough cell membrane and scattered piece-like growth. The growth of LST174/AS-EGFR cells was significantly as compared with that of the controls （optical density： 0.322 ±0.014 vs 0.422 ± 0.033, P 〈 0.05）, MTT assay showed that the inhibitory rate in LST174/AS-EGFR cells was lowered by 23.7% （P 〈 0.05）. In the experiment of colony formation on soft agar, LST174/AS-EGFR cells almost lost their abilities to form colony unlike the piled-up growth in the control cells.
CONCLUSION： AS-EGFR-induced blockage of endogenous EGFR self-secreted factor expression can not only inhibit the proliferation of colonic cancer cells, but also suppress the malignant transformation of phenotypes.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第35期3414-3416,共3页
World Chinese Journal of Digestology
关键词
表皮生长因子受体
反义寡聚核苷酸
结肠癌细胞系
细胞增殖
Epidermal growth factor receptor
Antisense oligonucleotide
Human colonic cancer cellline
Cell proliferation
