Chang Hong-min, Zhao Lei, Wang Xiao-jie, Fang Yan-hui, Li Dan, Luo Jing-mei, Du Luan-ying
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2014 Aug;32(4):299-303.
To investigate the effect of excretory/secretory proteins from Trichinella spiralis on apoptosis of NCI-H446 small-cell lung cancer cells.
Trichinella spiralis muscle stage larvae (5 x 10(6)/ml) were cultured in culture media for 24 h, the excretory/secretory proteins were collected from the supernatant of culture media. NCI-H446 small-cell lung cancer cells (No. A05) were randomly divided into three groups: experiment group (A), standard control group (apoptosis group, B), and control group (C). NCI-H446 cells in groups A and B were cultured with 0.3 mg/ml T. spiralis excretory/secretory proteins, and 6.4 microg/ml cisplatin for 24 h, respectively. NCI-H446 cells of group C were cultured for 24 h without any treatment. The expression of Bcl-2, Fas and Fasl mRNA was detected by RT-PCR. C-myc protein expression level was examined by Western blotting and immunofluorescence assay.
The level of Bcl-2 mRNA was lowest in group A(0.575 +/- 0.047) , Bcl-2 mRNA level in group C (0.975 +/- 0.069) was higher than that of group B (0.850 +/- 0.073) (P<0.05). Fas mRNA level was highest in group A (0.975 +/- 0.115), followed by group B (0.817 +/- 0.121) and group C(0.769 +/- 0.061) (P<0.05). The level of Fasl mRNA in groups A, B, and C was 0.669 +/- 0.051, 0.787 +/- 0.124, and 0.875 +/- 0.125, respectively (P<0.05). Fas/Fasl mRNA ratio in groups A, B, and C was 1.475, 1.038, and 0.878. Western blotting showed that the expression of C-myc protein in group C (1.172 +/- 0.026) was highest, followed by group B (1.074 +/- 0.069) and A (0.566 +/- 0.054) (P<0.05). Immunofluorescence test indicated that the C-mye protein was found in the cytoplasm and the nucleus 24 h after treated with 0.3 mg/ml T. spiralis excretory/secretory proteins and 6.4 p.g/ml cisplatin.
Trichinella spiralis excretory/secretory proteins may inhibit apoptosis of NCI-H446 small-cell lung cancer cells by reducing the apoptosis protein C-myc and Bcl-2 mRNA levels, and causing the increase of Fas/Fasl mRNA ratio.
探讨旋毛虫排泄/分泌蛋白对NCI-H446小细胞肺癌细胞凋亡的影响。
将旋毛虫肌幼虫（5×10⁶/ml）接种于培养基中培养24 h，收集培养基上清液中的排泄/分泌蛋白。将NCI-H446小细胞肺癌细胞（A05号）随机分为三组：实验组（A组）、标准对照组（凋亡组，B组）和对照组（C组）。A组和B组的NCI-H446细胞分别用0.3 mg/ml旋毛虫排泄/分泌蛋白和6.4 μg/ml顺铂培养24 h。C组的NCI-H446细胞未经任何处理培养24 h。采用RT-PCR检测Bcl-2、Fas和Fasl mRNA的表达。通过蛋白质印迹法和免疫荧光法检测C-myc蛋白表达水平。
A组Bcl-2 mRNA水平最低（0.575±0.047），C组（0.975±0.069）的Bcl-2 mRNA水平高于B组（0.850±0.073）（P<0.05）。A组Fas mRNA水平最高（0.975±0.115），其次为B组（0.817±0.121）和C组（0.769±0.061）（P<0.05）。A、B、C组Fasl mRNA水平分别为0.669±0.051、0.787±0.124和0.875±0.125（P<0.05）。A、B、C组Fas/Fasl mRNA比值分别为1.475、1.038和0.878。蛋白质印迹法显示，C组C-myc蛋白表达（1.172±0.026）最高，其次为B组（1.074±0.069）和A组（0.566±0.054）（P<0.05）。免疫荧光检测表明，用0.3 mg/ml旋毛虫排泄/分泌蛋白和6.4 μg/ml顺铂处理24 h后，C-mye蛋白在细胞质和细胞核中均有表达。
旋毛虫排泄/分泌蛋白可能通过降低凋亡蛋白C-myc和Bcl-2 mRNA水平，导致Fas/Fasl mRNA比值升高，从而抑制NCI-H446小细胞肺癌细胞凋亡。
