熊果酸对胰腺癌细胞SW1990增殖及其对微小RNA-760表达的影响

Effect of ursolic acid on the proliferation of pancreatic cancer cells SW1990 and the expression of its small RNA-760

目的探讨熊果酸对胰腺癌细胞SW1990增殖及其对微小RNA-760(miR-760)表达的影响。方法将SW1990细胞分为对照组和实验Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ组。对照组予以空白培养基溶液;实验Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ组分别予以含5,10,20,40,80μmol·L^(-1)熊果酸的培养基溶液培养。于作用24,48,72 h后,用四甲基偶氮唑盐(MTT)法测定细胞的增殖作用;用倒置显微镜观察细胞形态的变化;用microRNA提取试剂盒和反转录聚合酶链式反应测定miR-760的表达水平。结果对照组和实验Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ组的孵育24 h细胞存活率分别为(100.21±2.89)%,(94.42±2.24)%,(90.52±2.13)%,(86.71±0.48)%,(66.11±1.46)%,(40.50±1.32)%;孵育48 h细胞存活率分别为(100.38±2.56)%,(91.09±2.36)%,(85.54±7.43)%,(73.56±9.01)%,(52.72±1.25)%,(33.85±1.27)%;孵育72 h细胞存活率分别为(101.52±2.42)%,(84.66±2.51)%,(73.75±6.27)%,(63.10±2.08)%,(44.77±1.77)%,(24.43±1.30)%;孵育24 h miR-760表达量分别为1.00±0,1.15±0.23,1.47±0.41,1.77±0.27,2.65±0.14,3.54±0.51;孵育48h表达量分别为1.00±0,1.27±0.15,1.75±0.23,2.24±0.31,3.52±0.22,4.43±0.31;孵育72 h表达量别为1.00±0,1.40±0.11,2.00±0.10,2.52±0.17,3.93±0.65,5.16±0.45,差异均有统计学意义(均P<0.05)。随着熊果酸浓度增加和作用时间延长,SW1990细胞逐渐呈现变形、萎缩、甚至死亡。结论熊果酸对胰腺癌细胞具有显著增殖抑制作用,且呈浓度和时间依赖性,高浓度的熊果酸长时间作用可导致细胞miR-760表达量升高。

Objective To investigate effect of ursolic acid on the proli- feration of pancreatic cancer ceils 5W1990 and the expression of its small RNA-760 （miR- 760） . Methods The SW1990 cells were divided into control group and test Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ groups. Control group was given a blank medium solution; test Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ groups were givencuhured medium solution containing ursolic acid 5, 10, 20, 40, 80 μmol·L^-1 , respectively. After 24, 48, 72 h , the proliferation of ceils were measured by tetrazolium salt （MTT）; changes in cell morphology were observed by inverted microscope; the expression levels of miR -760 was determined by extraction kit and reverse transcription polymerase chain reaction. Results The cell survival rates of control group and test Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ groups after 24 h were （ 100. 21 ± 2. 89 ） %, （ 94. 42 ± 2.24 ） %, （ 90. 52 ± 2. 13 ） %, （86.71 ±0.48）% , （66. 11 ± 1.46）%, （40.50 ± 1.32）% , respectively; after 48 h, cell survival rates were （100.38±2.56）%, （91.09 ±2.36）%, （85.54 ±7.43）%, （73.56 ±9.01）%, （52.72 ± 1.25）%, （33.85 ± 1.27） %, respectively; after 72 h, cell survival rates were （ 101.52 ± 2. 42 ） %, （ 84. 66 ± 2.51 ） %, （73.75 ±6.27）%, （63.10 ±2.08）%, （44.77 ±1.77）%, （24.43 ±1.30）% respectively; after 24 h, the expression of miR - 760 were 1.00 ±0, 1.15 ±0.23, 1.47 ±0.41, 1.77 ±0.27, 2.65 ±0.14, 3.54 ±0.51, respectively; after48 h, the of expression miR-760 were 1.00 ±0, 1.27 ±0.15, 1.75 ±0.23, 2.24 ±0.31, 3.52 ±0. 22, 4.43 ± 0. 31, respectively; after 72 h, the expression of miR - 760 werel. 00 ± 0, 1.40 ± 0. 11, 2. 00 ± 0. 10, 2.52 ± 0. 17, 3.93 ± 0. 65, 5.16 ± 0. 45, respectively, ursolic acid groups had statistically significant differences compared with the control group （ P 〈 0.05 ）. With the increase of the concentration of ursolie acid and the prolongation of action time, SW1990 cells gradually exhibit deformation, shrink, or even death, miR - 760 levels increased. Conelusion Ursolic acid has a significant value - added inhibition on the pancreatic cancer cells in a dose and time dependent, and a high concentration of ursolic acid under a prolonged action time may result in an increased expression of miR - 760 cells.