摘要
目的评价异氟醚对人舌癌细胞（Tea8113细胞）基质金属蛋白酶．2表达的影响。方法取对数生长期的Tea8113细胞，制成单细胞悬液，接种于96孔培养板（5×10^3／ml，200μl/孔）、6孔培养板（1×105／ml，5ml／孔）、培养皿（5×10^5／ml，5ml／皿）和Transwell小室[上层小室加入细胞悬液200μl（细胞密度5×10^5／ml），下层小室加入10％血清培养基500μl]，常规培养待细胞完全贴壁后进行实验。采用随机数字表法，将细胞分为3组（n=30）：对照组（C组）、2％异氟醚2h组（I1组）、2％异氟醚4h组（12组）。C组不给予异氟醚，I1组和12组细胞培养液中加入2％异氟醚分别孵育细胞2或4h。采用MTT法检测细胞活力，流式细胞术检测细胞凋亡率；以细胞划痕实验及Transwell小室实验检测细胞水平迁移和侵袭能力；采用免疫细胞化学及RT-PCR检测细胞MMP-2及其mRNA表达水平。结果与C组比较，I1组、12组细胞活力和细胞水平迁移率升高，细胞凋亡率降低，MMP-2及其mRNA表达上调，I1组透膜细胞数增多（P〈0．05）。与I1组比较，I2组细胞活力和细胞水平迁移率升高，细胞凋亡率降低，透膜细胞数增多，MMP-2及其mRNA表达上调（P〈0．05）。结论异氟醚可能通过上调人舌癌Tca8113细胞的MMP-2表达来促进癌细胞的迁移侵袭能力。
Objective To evaluate the effect of isoflurane on the expression of matrix metalloproteinase 2 （MMP-2） in human tongue cancer cell. Methods Tea8113 cells at the logarithmic growth phase were seeded into 96-well plates （200 /d/hole） with the density of 5 × 10^3/ml, 6-well plates （5 ml/hole） with the density of 1 ×10^5/ml, culture dishes （5 ml/dish） with the density of 5 × 10^5/ml or Transwell chamber （200 p.l cell suspension for upper chamber with a density of 5 × 10^5/ml, 10% blood serum medium 500 μl for lower chamber）. The cells were randomly divided into 3 group （n = 30 each） ： control group （group C）, 2% isoflurane 2 h group （group I1 ） and 2% isuflurane 4h group （group I2 ）. 2% isoflurane was added to the culture medium and the cells were incubated for 2 and 4 h in groups I1 and I2, respectively. The cell viability was detected by MTT assay. The cell apoptosis was assessed by flow cytometry. The migration and invasion of the cells were measured by cell wound scratch assay and Transwell chamber assay, respectively. The expression of MMP-2 and MMP-2 mRNA was detected by immu- nocytochemistry and RT-PCR, respectively. Results The cell viability and migration of cells were significantly in- creased, the apoptotic rate was decreased, and the expression of MMP-2 and MMP-2 mRNA was up-regnlated in groups 11 and I2, and the invasion of the cells were increased in group 12 as compared with group C （ P 〈 0.05）. Compared with group I1, the cell viability and migration and invasion of the cells were significantly increased, the apoptotic rate was decreased, and the expression of MMP-2 and MMP-2 mRNA was up-regulated in group I2 （ P 〈0.05）. Conclusion Isoflurane enhances the migration and invasion of the cancer cells by up-regulating the ex- pression of MMP-2 in human tongue cancer Tea8113 cells.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2013年第3期317-319,共3页
Chinese Journal of Anesthesiology
关键词
异氟醚
舌肿瘤
细胞系
肿瘤
基质金属蛋白酶2
Isoflurane
Tongue neoplasms
Cell line, tumor
Matrix metalloproteinase 2
