摘要
目的 观察改变胞内MnSOD基因表达水平对SGC7901胃癌细胞增殖的影响。方法 采用电穿孔方法将反义和正义MhSOD cDNA真核表达载体pHβA-SOD(-)/pHβA-SOD(+)转染SGC7901胃癌细胞,400mg/LG418筛选稳定表达克隆并用RT-PCR及Western杂交法鉴定后扩大培养。用pHβA空质粒转染作为对照。放射免疫法检测正义、反义及空载SGC7901胃癌细胞株内的铜锌超氧化物歧化酶(CuZnSOD,SOD1)蛋白含量。分别用直接细胞计数法、四唑蓝(MTT)比色法、平皿克隆形成率试验及裸鼠移植瘤模型测定3组细胞在体外、体内的增殖活性。结果 与空载组(Vector—7901)相比,转染正义质粒的MnSOD-7901胃癌细胞呈现抑增殖效应:(1)生长曲线示增殖速度减慢;(2)在平皿上的集落形成能力下降;(3)在裸鼠体内的成瘤性明显受抑制。转染反义质粒的MnSOD-AS7901胃癌细胞则出现促增殖效应:(1)生长曲线示增殖速度加快;(2)在平皿上的集落形成率上升;(3)裸鼠移植瘤生长加速。结论 通过基因转染提高胞内MnSOD的表达可抑制胃癌细胞的增殖,MnSOD可能是胃癌基因治疗的一个新靶点。
Purpose: To observe the effects on proliferation of SGC7901 mediated by MnSOD gene transfection. Methods: We transferred sense and antisense MnSOD cDNA carrier pHβA-SOD (-)/pHβA-SOD (+) or mock vector into SGC7901 gastric cancer cell by electroporation. Then we screened stable expression sublines by 400 μg/mL G418 solution and further confirmed by RT-PCR and Western blot. Then the subtypes of SGC7901 were marked as MnSOD-7901 MnSOD-AS7901 and vector-7901, respectively. The Copper-Zinc superoxide dismutase (CuZnSOD) contents in the transfectants were also measured by radioimmunoassay. We also evaluated the proliferation activities in these three subtypes by direct cells count, MTT assay, plate clone formation test and nude mice transplanting tumor model, respectively. Results: Compared with the vector-7901, MnSOD-74901 revealed such characteristics as followed: (a) Partial inhibition of proliferation activities reflected by growth curves; (b) Decreasing clone formation on plate; (c) Remarkable growth retardation of transplanting tumor in nude mice. On the contrary, the MnSOD-AS7901 subline revealed the promotion of tumor proliferation, which was indicated as followed: (a) Promoting the proliferation activities under cells culture; (b) Increasing clone formation on plate; (c) Remarkable growth acceleration of nude mice tumor transplantation model. Conclusions: MnSOD transfection can inhibit the growth of SGC7901 gastric cancer cell. Thus MnSOD can be a novel target for gene therapy.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
2004年第1期19-23,31,共6页
Fudan University Journal of Medical Sciences
基金
国家自然科学基金(39770330)
