Department of Gastroenterology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, PR China.
J Exp Clin Cancer Res. 2011 Jul 7;30(1):67. doi: 10.1186/1756-9966-30-67.
Our study aims to evaluate the anti-growth effects of recombinant immunotoxin (IT) anti-c-Met/PE38KDEL on gastric cancer cells, and its mechnisms.
Gastric cancer cells were treated with increasing doses of IT and c-Met protein was quantified by Western blotting. Cell proliferation was determined by Cell Counting Kit-8 assay (CCK). [3H]-leucine incorporation assay was used to evaluate IT inhibition of protein synthesis. Cell apoptosis was quantified by flow cytometry. Caspase activities were measured using colorimetric protease assays.
Cell growth and protein synthesis of the gastric cancer cell lines were suppressed by IT in a dose- and time-dependent manner. IT also induced apoptosis in a dose-dependent manner. The apoptosis rates of gastric cancer cell lines MKN-45 and SGC7901 were 19.19% and 27.37%, respectively when treated with 50 ng/ml of IT. There were significant increase of caspase-3 activity at 24 hr of IT treatment (100 ng/ml) (P < 0.01) in these gastric cancer cell lines.
IT anti-c-Met/PE38KDEL has anti-growth effects on the gastric cancer cell lines in vitro, and it provides an experimental basis for c-Met-targeted therapy towards in vivo testing.
本研究旨在评估重组免疫毒素（IT）抗 c-Met/PE38KDEL 对胃癌细胞的抗生长作用及其机制。
用递增剂量的 IT 处理胃癌细胞，并用 Western blot 定量 c-Met 蛋白。用细胞计数试剂盒-8 法（CCK）测定细胞增殖。[3H]-亮氨酸掺入法评估 IT 对蛋白质合成的抑制作用。用流式细胞术定量细胞凋亡。用比色蛋白酶测定法测定半胱天冬酶活性。
IT 以剂量和时间依赖的方式抑制胃癌细胞系的细胞生长和蛋白质合成。IT 还以剂量依赖的方式诱导细胞凋亡。当用 50ng/ml 的 IT 处理时，胃癌细胞系 MKN-45 和 SGC7901 的凋亡率分别为 19.19%和 27.37%。用 IT 处理 24 小时后，这些胃癌细胞系中的 caspase-3 活性显著增加（100ng/ml）（P<0.01）。
IT 抗 c-Met/PE38KDEL 对体外胃癌细胞系具有抗生长作用，为 c-Met 靶向治疗提供了体内试验的实验基础。
