| 赵舰,张良龙,金亮,等.白消安对U87胶质瘤干细胞增殖和自我更新的影响[J].安徽医药,2020,24(9):1787-1790. |
| 白消安对U87胶质瘤干细胞增殖和自我更新的影响 |
| Effect of Busulfan on proliferation and self-renewal of U87 glioma stem cells |
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| DOI：10.3969/j.issn.1009-6469.2020.09.023 |
| 中文关键词: 神经胶质瘤 细胞分裂 细胞凋亡 半胱氨酸天冬氨酸蛋白酶 3 bcl-2相关 X蛋白质 细胞增殖 白消安 U87胶质瘤干细胞 |
| 英文关键词: Glioma Cell division Apoptosis Caspase 3 Bcl-2-associated X protein Cell proliferation Busulfan U87 glioma stem cells |
| 基金项目:河北省沧州市重点研发计划指导项目( 183302129) |
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| 中文摘要: |
| 目的探究白消安对 U87胶质瘤干细胞增殖和自我更新的影响。方法培养 U87胶质瘤干细胞，分为四组， A组加入 200 μL含 5%胎牛血清的 DMEM/F12培养基， B、C、D三组分别加入 200 μL含 20、50、100 μmol/L白消安的 DMEM/F12培养基。比较四组细胞增殖情况［四甲基偶氮唑盐微量酶反应比色法( MTT法)］、细胞凋亡情况［即 B细胞淋巴瘤 /白血病 -2(Bcl-2)、Bcl-2相关 X蛋白( Bax)、半胱氨酸天冬氨酸蛋白酶 3(Caspase-3)表达水平］，以及细胞周期。结果培养 24 h后，四组 U87胶质瘤干细胞吸光度比较差异无统计学意义( P＞0.05)；培养 48 h、72 h后， B、C、D组 U87胶质瘤干细胞吸光度低于 A组( P＜0.05)；培养 48 h、72 h后， B、C、D组 U87胶质瘤干细胞吸光度比较差异无统计学意义( P＞0.05)。 C组 U87胶质瘤干细胞 Bax、Caspase-3、 Bax/Bcl-2的吸光度值、阳性细胞表达率高于 A、B组，比较差异有统计学意义［吸光度值: CBax(0.38±0.01)比 ABax(0.46±0.04)、 BBax(0.41±0.01)； CCaspase-3(0.45±0.02)比 ACaspase-3(0.54±0.01)、 BCaspase-3(0.47±0.02)； CBax/Bcl-2(1.19±0.04)比 ABax/Bcl-2(1.36±0.01)、 BBax/Bcl-2(1.22±0.04)；阳性细胞表达率: CBax(47.53±1.34)%比 ABax(39.81±4.02)%、BBax(39.51±1.37)%；CCaspase-3(65.17±2.34)%比 ACaspase-3(54.42±1.34)%、BCaspase-3(49.16±2.31)%；CBax/Bcl-2(60.24±4.10)%比 ABax/Bcl-2(35.47±1.20)%、BBax/Bcl-2(37.41±4.12)%；P＜0.05］； D组 U87胶质瘤干细胞 Caspase-3、Bax/Bcl-2的吸光度值、阳性细胞表达率高于 A、B组，比较差异有统计学意义［吸光度值: DBax(0.37±0.01)比 ABax(0.46±0.04)、 BBax(0.41±0.01)； DCaspase-3(0.28±0.01)比 ACaspase-3(0.54±0.01)、 BCaspase-3(0.47±0.02)； DBax/Bcl-2(0.97±0.02)比 ABax/Bcl-2(1.36±0.01)、 BBax/Bcl-2(1.22±0.04)；阳性细胞表达率: DCaspase-3(68.06±1.09)%比 ACaspase-3(54.42±1.34)%、BCaspase-3(49.16±2.31)%；DBax/Bcl-2(65.10±2.15)%比 ABax/Bcl-2(35.47±1.20)%、BBax/Bcl-2(37.41±4.12)%；P＜0.05］。 A组、 D组处于 G1期细胞分别占8.20%、3.22%，处于 G2 /M期细胞分别占 16.07%、12.18%，两组比较差异有统计学意义( P＜0.05)； A组、 D组处于 G0/G1期细胞分别占 52.27%、58.85%，处于 S期细胞分别占 23.36%、35.75%，两组比较差异无统计学意义( P＞0.05)。结论白消安可抑制 U87胶质瘤干细胞的增殖，促进其凋亡，并且呈现一定量效关系；机制可能与白消安上调 Bcl-2、Bax、Caspase-3表达有关。 |
| 英文摘要: |
| Objective To explore the effect of Busulfan on proliferation and self-renewal of U87 glioma stem cells.Methods U87 glioma stem cells were cultured and divided into four groups.Group A was cultured with 200 μL of DMEM/F12 medium containing5% fetal bovine serum.Groups B，C，and D were cultrued with 200 μL of DMEM/F12 medium containing 20，50，and 100 μmol/L busulfan，respectively.The cell proliferation of the four groups［tetramethyl azozole trace enzyme reaction colorimetric method(MTT method)］，apoptosis［ie.B cell lymphoma/leukemia-2(Bcl-2)Bcl-2 related X protein(Bax)caspase-3 expression levels］，and the cell cycle were compared.Results After 24 hours of culture，t，here was no significant difference in the absorbance of U87 glioma stem cells among the four groups(P＞0.05)； after 48 hours and 72 hours of culture，the absorbance of U87 glioma stem cells in groups B，C，and D was lower than that in group A(P＜0.05).After 48 h and 72 h incubation，there was no significant difference in the absorbance of U87 glioma stem cells in groups B，C and D(P＞0.05).The absorbance value and positive expression rate of Bax，Caspase-3，Bax/Bcl-2 of U87 glioma stem cells in group C were higher than those in groups A and B，and the differences were statistically significant(absorbance value:CBax(0.38±0.01)vs. ABax(0.46±0.04)，BBax(0.41±0.01)； CCaspase-3(0.45±0.02)vs. ACaspase-3(0.54±0.01)BCaspase-3(0.47±0.02)； CBax/Bcl-2(1.19± 0.04)vs. ABax/Bcl-2(1.36±0.01)BBax/Bcl-2(1.22±0.04)； positive cell expression rate: CBax(47.53±14)% vs. ABax(39.81±4.02)%，BBax(39.51 ±1.37)%；CCaspase-3(6517±2.34)% vs. ACaspase-3(54.42±1.34)%，BCaspase-3.3(49.16±2.31)%；CBax/Bcl-2(60.24±4.10)% vs. ABax/Bcl-2(35.47±1.20)%，BBax/Bcl-2(37.41±4.12)%；P＜0.05)； absorbance value of caspase-3，Bax/Bcl-2，positive cell expression rate of U87 glioma stem cells in group D were higher than those in group A and B，the differ- ences were statistically significant(absorbance value:DBax(0.37±0.01)vs. ABax(0.46±0.04)，BBax(0.41±0.01)； DCaspase-3(0.28±0.01) vs. ACaspase-3(0.54±0.01)，BCaspase-3(0.47±0.02)； DBax/Bcl-2(0.97±0.02)vs. ABax/Bcl-2(1.36±0.01)，BBax/Bcl-2(1.22±0.04)； positive Cell ex- pression rate:DCaspase-3(68.06±1.09)% vs. ACaspase-3(54.42±1.34)%，BCaspase-3(49.16±2.31)%；DBax/Bcl-2(65.10±2.15)% vs. ABax/Bcl-2(35.47±1.20)%，BBax/Bcl-2(37.41±4.12)%；P＜0.05).In group A and D，cells in G1 phase accounted for 8.20% and 3.22%，respectively，，and cells in G2/M accounted for 16.07% and 12.18%，respectively.The difference between the two groups was statistically significant(P＜0.05).In group A and D，cells in the G0/G1 phase accounted for 52.27% and 58.85%，and the cells in the S phase accounted for 23.36% and 35.75%，respectively.There was no statistically significant difference between the two groups(P＞0.05).Conclusion Busulfan can inhibit the proliferation of U87 glioma stem cells，promote their apoptosis，and show a dose-effect relationship.The mechanism may be related to Busulfan’s up-regulation of Bcl-2，Bax，Caspase-3 expression. |
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