Zhao Jiaying, Cai Yuankun, Yin Chenqing, Lv You, Wei Wanmin, Wang Xin, Hao Zong, Shen Chenxia, Wang Huipeng, Chen Jun
Department of General Surgery, The Fifth People's Hospital of Shanghai, Fudan University, Shanghai 200240, P.R. China.
Central Laboratory, The Fifth People's Hospital of Shanghai, Fudan University, Shanghai 200240, P.R. China.
Oncol Rep. 2016 Feb;35(2):985-91. doi: 10.3892/or.2015.4446. Epub 2015 Nov 25.
The present study explored the inhibitory effect of hyperthermic CO2 pneumoperitoneum on the proliferation and migration of colon cancer cells, and its mechanism. Colon cancer cell line SW-480 was sealed into a urine collection bag to simulate pneumoperitoneum with 100% CO2 under a pressure of 14 mmHg. The growth and morphology of cells were observed under a microscope, the inhibition on cell proliferation was measured using WST-8 test, cell apoptosis and the cell cycle were monitored using fluorescence-activated cell sorting analysis, the migration of cells was tested using the scratch assay, and the expression of HSP-70, caspase-3, hypoxia-inducible factor-1α (HIF-1α) and matrix metalloproteinase-9 (MMP-9) proteins and genes was investigated using western blotting and reverse transcription polymerase chain reaction. Compared with the control group, there was no significant difference in the CO2 group (P>0.05), while the apoptosis and necrosis rates in the hyperthermo-CO2 group was significantly increased (P<0.05). Compared with the control group, the number of cells at G0/G1 phase significantly increased and the number of cells at S phase significantly decreased in the hyperthermo-CO2 group (P<0.05), indicating that hyperthermo-CO2 could arrest the cell cycle. It was suggested by the results of the scratch assay that cell migration ability enhanced in the CO2 group, but decreased in the hyperthermo-CO2 group compared with the control. CO2 pneumoperitoneum promoted cell migration by upregulating HIF-1α and MMP-9 expression. However, the CO2 pneumoperitoneum with hyperthermia enhanced apoptosis and inhibited migration by upregulating the expression of HSP-70, HIF-1α and caspase-3, but downregulating the expression of MMP-9.
本研究探讨了高温二氧化碳气腹对结肠癌细胞增殖和迁移的抑制作用及其机制。将结肠癌细胞系SW-480密封于尿液收集袋中，在14 mmHg压力下用100%二氧化碳模拟气腹。在显微镜下观察细胞的生长和形态，采用WST-8法检测对细胞增殖的抑制作用，采用荧光激活细胞分选分析监测细胞凋亡和细胞周期，采用划痕试验检测细胞迁移，并采用蛋白质印迹法和逆转录聚合酶链反应研究热休克蛋白70（HSP-70）、半胱天冬酶-3（caspase-3）、缺氧诱导因子-1α（HIF-1α）和基质金属蛋白酶-9（MMP-9）蛋白及基因的表达。与对照组相比，二氧化碳组差异无统计学意义（P>0.05），而热二氧化碳组的凋亡和坏死率显著升高（P<0.05）。与对照组相比，热二氧化碳组G0/G1期细胞数量显著增加，S期细胞数量显著减少（P<0.05），表明热二氧化碳可使细胞周期停滞。划痕试验结果表明，与对照组相比，二氧化碳组细胞迁移能力增强，而热二氧化碳组细胞迁移能力降低。二氧化碳气腹通过上调HIF-1α和MMP-9表达促进细胞迁移。然而，高温二氧化碳气腹通过上调HSP-70、HIF-1α和caspase-3的表达，但下调MMP-9的表达，增强细胞凋亡并抑制细胞迁移。
