高压氧联合顺铂对肺腺癌A549／DDP细胞增殖的影响

In vitro effects of hyperbaric oxygen and DDP on proliferation of lung adenocarcinoma cell line A.549/DDP

目的观察高压氧（HBO）联合顺铂（DDP）对肺腺癌A549／DDP细胞增殖的影响以及高压氧对细胞周期和凋亡的影响。方法采用四甲基偶氮唑蓝（MTT）法检测高压氧和顺铂对A549／DDP细胞是否有协同抑制作点H，流式细胞法（FCM）分析高压氧对细胞周期和细胞凋亡的影响。结果单纯高压氧对A549／DDP细胞的抑制率为2．90％，与常压对照组比较差异有统计学意义（P〈0．05）。不同浓度（3、6、12、24μg／m1）的顺铂对A549／DDP细胞的抑制率分别为1．87％、4．62％、11．15％、30．45％，各组间比较差异均有显著性（P〈0．01）；高压氧联合不同浓度顺铂对A549／DDP细胞的抑制率分别为4．37％、8．92％、21．88％、48．71％，与顺铂组比较差异均有统计学意义（P〈0．05）。FCM结果显示，A549／DDP细胞经高压氧干预后，GO^G1期细胞减少，s期细胞增多，与对照组比较差异均有极显著性（P〈0．01）。凋亡率由2．8％增至7．9％，差异有统计学意义（P〈0．01）。结论高压氧与顺铂可起到协同抑制A549／DDP细胞增殖的作用，高压氧能阻滞细胞于s期，并促进细胞凋亡。

Objective To evaluate the effects of hyperbaric oxygen（HBO） and cisplatin（DDP） on inhibition of A549/DDP cells in vitro and to explore the impacts of HBO on cell cycle and apoptosis. Methods The cytotoxicity of HBO and DDP was determined by Methyl thiazolyl tetrazolium （MrI＇F） assay; flow cytometry （FCM） was used to investigate the impacts on cell cycle and apoptosis of cell line A549/DDP. Results Hyperbaric oxygen alone significantly suppressed A549/ DDP ceils proliferation（P 〈0.05） , compared with control group of normal oxygen, and the inhibition ratio was 2.90%. DDP at concentrations ranging from 3 μg/ml to 24 μg/ml significantly inhibited the proliferation of A549/DDP cells in a dose dependent manner, that was concentrations of 3,6, 12 and 24 μg/ml brought inhibition ratio of 1.87% , 4.62% , 11.15% and 30. 45 % ; Hyperbaric oxygen can enhance the inhibition effect of DDP to A549/DDP cells, when hyperbaric oxygen affiliated with DDP of concentrations above, the inhibition ratios were 4. 37%, 8.92%, 21.88% and 48.71%, respectively, which has significant difference with the single DDP groups. FCM analyses showed that HBO arrested A549/DDP cells at the S phase, that was cell number of G0/GI phase reduced significantly and of S phase raised significantly, compared with normal oxygen group （ P 〈 0.01 ） , that was the percentages of cells in S phase of control group were 9.5 % and HBO group were 20.9% , while the percentages of ceils in GO/Glphase were 82.2% and 66.3% , respectively. In addition, there were more apoptotic cells in the HBO group with the percentage of 7.9%, while the percentage was 2.8% in the normal oxygen group. Conclusion Hyperbaric oxygen could play together with DDP to inhibit A549/DDP cells proliferation. HBO could block the cell cycle at S period and induce apoptosis.