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目的:体外模拟腹腔镜手术CO2气腹方法,研究不同压力CO2环境对人宫颈癌Hela细胞黏附和侵袭能力的影响。方法:培养人宫颈癌Hela细胞株,模拟CO2气腹压力12 mmHg、16 mmHg处理4 h,以培养于常规条件下的人宫颈癌Hela细胞为对照,通过MTT法检测细胞活性,通过细胞基质黏附试验检测细胞基质黏附能力改变,应用免疫组织化学方法检测人宫颈癌Hela细胞中E-钙黏素(E-cadherin)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达变化。结果:与对照组相比,CO2气腹处理组生长曲线上移(P<0.05)、随压力增加生长速度加快;气腹组与对照组相比,细胞基质黏附能力增强(P<0.05)、E-钙黏素表达减少(P<0.05)、MMP-9表达增加(P<0.05)。但不同压力条件下E-钙黏素和MMP-9表达水平无统计学差异(P>0.05),细胞基质黏附能力无明显增加。结论:CO2气腹环境增加了宫颈癌的侵袭和转移的风险,其机制可能与人宫颈癌Hela细胞的E-钙黏素的表达降低,MMP-9的表达增加有关,不同压力的CO2气腹对宫颈癌的侵袭和转移能力无明显影响。
OBJECTIVE: To simulate CO2 pneumoperitoneum in laparoscopic surgery in vitro to study the effect of CO2 environment on the adhesion and invasion of human cervical carcinoma Hela cells. Methods: The human cervical cancer Hela cell line was cultured and simulated by CO2 pneumoperitoneum pressure 12 mmHg and 16 mmHg for 4 h. The cultured human cervical cancer Hela cells under normal conditions were used as control. The cell viability was detected by MTT assay. The changes of cell adhesion were observed by immunohistochemistry. The expression of E-cadherin and matrix metalloproteinase-9 (MMP-9) in human cervical carcinoma Hela cells was detected by immunohistochemistry. Results: Compared with the control group, the growth curve of CO2 pneumoperitoneum group was up-shifted (P <0.05), and the growth rate was accelerated with the increase of pressure. Compared with the control group, the pneumoperitoneum group showed enhanced cell adhesion (P <0.05) E-cadherin decreased (P <0.05), and MMP-9 increased (P <0.05). However, there was no significant difference in E-cadherin and MMP-9 expression under different stress conditions (P> 0.05), but there was no significant increase in cell adhesion. CONCLUSION: CO2 pneumoperitoneum increases the risk of cervical cancer invasion and metastasis. The mechanism may be related to the decrease of E-cadherin expression and MMP-9 expression in human cervical cancer Hela cells. Different pressures of CO2 pneumoperitoneum Cervical cancer invasion and metastasis had no significant effect.