[1]刘胜,赵梦圆,金巍,等.对映-贝壳杉烯型二萜类化合物J32对胰腺癌细胞的作用及机制[J].新乡医学院学报,2023,40(9):810-817.[doi:10.7683/xxyxyxb.2023.09.002]
　LIU Sheng,ZHAO Mengyuan,JIN Wei,et al.Effect and mechanism of ent-kaurene diterpenoid J32 on pancreatic cancer cells[J].Journal of Xinxiang Medical University,2023,40(9):810-817.[doi:10.7683/xxyxyxb.2023.09.002]
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对映-贝壳杉烯型二萜类化合物J32对胰腺癌细胞的作用及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
40卷
期数:
2023年9
页码:
810-817
栏目:
基础研究
出版日期:
2023-09-05

文章信息/Info

Title:
Effect and mechanism of ent-kaurene diterpenoid J32 on pancreatic cancer cells
作者:
刘胜1; 赵梦圆2; 金巍1; 段长恩1; 可钰3
（1.新乡医学院第一附属医院普通外科，河南　卫辉　453100;2.河南医药健康技师学院，河南　开封　475006;3.郑州大学药学院，河南　郑州　450001）
Author(s):
LIU Sheng1; ZHAO Mengyuan2; JIN Wei1; DUAN Changen1; KE Yu3
(1.Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;2.Henan Medical and Health Technician College,Kaifeng 475006,Henan Province,China；3.College of Pharmacy,Zhengzhou University,Zhengzhou 450001,Henan Province,China)
关键词:
对映-贝壳杉烯型二萜; DNA损伤; 增殖; 迁移; 凋亡; 活性氧; 共济失调毛细血管扩张和Rad3相关蛋白-细胞周期检查点激酶1-p53-Cyclin D1通路
Keywords:
ent-kaurane diterpenoid; DNA damage; proliferation; migration; apoptosis; reactive oxygen species; ataxia telangiectasia and Rad3 related protein-checkpoint kinase 1-p53-Cyclin D1 pathway
分类号:
R735.9
DOI:
10.7683/xxyxyxb.2023.09.002
文献标志码:
A
摘要:
目的　探讨对映-贝壳杉烯型二萜类化合物 J32对胰腺癌细胞SW1990的作用及机制。
方法　将SW1990细胞随机分为对照组、0.5 μmol·L-1 J32组、1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组、4.0 μmol·L-1 J32组，分别用含终浓度0.0、0.5、1.0、2.0、4.0 μmol·L-1 J32的培养基培养。采用细胞划痕试验检测细胞迁移能力，单克隆形成试验检测细胞增殖能力，流式细胞术检测细胞凋亡情况、细胞中活性氧水平及细胞周期，磷酸化H2组蛋白家族成员X（γ-H2AX）免疫荧光法检测细胞DNA损伤程度，Western blot法检测共济失调毛细血管扩张和Rad3相关蛋白（ATR）-细胞周期检查点激酶1（Chk1）-p53-细胞周期素D1（Cyclin D1）通路相关蛋白以及凋亡相关蛋白B细胞淋巴瘤-2相关X蛋白(Bax)、caspase-3、B淋巴细胞瘤-2（Bcl-2）表达。
结果　培养24、48 h 时，1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组、4.0 μmol·L-1 J32组细胞的迁移能力显著低于对照组(P<0.05)，2.0 μmol·L-1 J32组、4.0 μmol·L-1 J32组细胞的迁移能力均显著低于1.0 μmol·L-1 J32组(P<0.05)，4.0 μmol·L-1 J32组细胞的迁移能力显著低于2.0 μmol·L-1 J32组 (P<0.05) 。0.5 μmol·L-1 J32组、1.0 μmol·L-1 J32组细胞的增殖能力显著低于对照组(P<0.05)，1.0 μmol·L-1 J32组细胞的增殖能力显著低于 0.5 μmol·L-1 J32组 (P<0.05)。1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组、 4.0 μmol·L-1 J32组细胞的凋亡率显著高于对照组(P<0.05)，2.0 μmol·L-1 J32组、 4.0 μmol·L-1 J32组细胞的凋亡率显著高于1.0 μmol·L-1 J32组(P<0.05)，4.0 μmol·L-1 J32组细胞的凋亡率显著高于2.0 μmol·L-1 J32组(P<0.05)。1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组、 4.0 μmol·L-1 J32组细胞的活性氧水平显著高于对照组(P<0.05)，2.0 μmol·L-1 J32组、4.0 μmol·L-1 J32组细胞的活性氧水平显著高于1.0 μmol·L-1 J32组(P<0.05)，4.0 μmol·L-1 J32组细胞的活性氧水平显著高于 2.0 μmol·L-1 J32组 (P<0.05)。 1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组细胞的DNA损伤程度显著高于对照组(P<0.05)，2.0 μmol·L-1 J32组细胞的DNA损伤程度显著高于1.0 μmol·L-1 J32组 (P<0.05)。1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组细胞中G1/S期细胞占比显著高于对照组(P<0.05)，2.0 μmol·L-1 J32组细胞中G1/S期细胞占比显著高于1.0 μmol·L-1 J32组 (P<0.05)。对照组、1.0 μmol·L-1 J32组、2.0 μmol·L-1 J32组、4.0 μmol·L-1 J32组细胞中，随着J32浓度的增加，磷酸化ATR/ATR值及磷酸化p53蛋白表达呈升高趋势（F=25.534、3.970，P<0.05），Chk1、CyclinD1蛋白表达呈降低趋势（F=19.532、0.485，P<0.05）；促凋亡蛋白caspase-3、Bax的表达呈升高趋势（F=0.219、4.314，P<0.05），抗凋亡蛋白Bcl-2表达呈下降趋势（F=0.324，P<0.05）。
结论　对映-贝壳杉烯型二萜类化合物J32可呈浓度依赖性地抑制胰腺癌SW1990细胞的迁移和增殖、促进细胞凋亡及提高细胞内活性氧水平，此外，J32可促进细胞发生DNA损伤及细胞周期阻滞，其作用机制可能与ATR-Chk1-p53-Cyclin D1通路相关。
Abstract:
Objective　To investigate the effect and mechanism of ent-kaurene diterpenoid J32 on pancreatic cancer cell SW1990.
Methods　The SW1990 cells were randomly divide into the control group,0.5 μmol·L-1 J32 group,1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group,and were cultured with medium containing final concentrations of 0.0,0.5,1.0,2.0 and 4.0 μmol·L-1 J32,respectively.The cell migration ability was detected by cell scratch assay;the cell proliferation ability was detected by monoclonal formation assay;the cell apoptosis,reactive oxygen species level in cells and cell cycle were detected by flow cytometry;the degree of cellular DNA damage was detected by phosphorylated H2A histone family member X (γ-H2AX） immunofluorescence assay;the expressions of ataxia telangiectasia and Rad3 related protein (ATR）-checkpoint kinase 1 (Chk1）-p53-CyclinD1 pathway related proteins,apoptosis related proteins B-cell lymphoma-2 associated X protein (Bax),caspase-3,B-cell lymphoma-2 (Bcl-2）were detected by Western blot.
Results　At 24 and 48 hours of cultivation,the migration abilities of cells in the 1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly lower than those in the control group (P<0.05);the migration abilities of cells in the 2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly lower than those in the 1.0 μmol·L-1 J32 group (P<0.05);the migration ability of cells in the 4.0 μmol·L-1 J32 group was significantly lower than that in the 2.0 μmol·L-1 J32 group (P<0.05).The proliferation ability of cells in the 0.5 μmol·L-1 J32 group,1.0 μmol·L-1 J32 group were significantly lower than those in the control group (P<0.05);the proliferation ability of cells in the 1.0 μmol·L-1 J32 group was significantly lower than that in the 0.5 μmol·L-1 J32 group(P<0.05).The cell apoptosis rates of cells in the 1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly higher than those in the control group (P<0.05);the cell apoptosis rates of cells in the 2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly higher than those in the 1.0 μmol·L-1 J32 group (P<0.05);the cell apoptosis rate of cells in the 4.0 μmol·L-1 J32 group was significantly higher than that in the 2.0 μmol·L-1 J32 group (P<0.05).The reactive oxygen species levels of cells in the 1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly higher than those in the control group (P<0.05);the reactive oxygen species levels of cells in the 2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group were significantly higher than those in the 1.0 μmol·L-1 J32 group (P<0.05);the reactive oxygen species level of cells in the 4.0 μmol·L-1 J32 group was significantly higher than that in the 2.0 μmol·L-1 J32 group (P<0.05).The degree of DNA damage of cells in the 1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group was significantly higher than those in the control group (P<0.05);the degree of DNA damage of cells in the 2.0 μmol·L-1 J32 group was significantly higher than that in the 1.0 μmol·L-1 J32 group (P<0.05).The proportions of G1/S phase cells in the 1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group were significantly higher than those in the control group (P<0.05);the proportion of G1/S phase cells in the 2.0 μmol·L-1 J32 group was significantly higher than that in the 1.0 μmol·L-1 J32 group (P<0.05).In the control group,0.5 μmol·L-1 J32 group,1.0 μmol·L-1 J32 group,2.0 μmol·L-1 J32 group,4.0 μmol·L-1 J32 group,with the increase of J32 concentration,the phospho-ATR/ATR value and phospho-p53 protein expression showed increasing trend (F=25.534,3.970; P<0.05),while the expression of Chk1 and CyclinD1 proteins showed decreasing trend (F=19.532,0.485;P<0.05);the expression of pro-apoptotic proteins caspase-3 and Bax showed increasing trend (F=0.219,4.314；P<0.05),while the expression of anti-apoptotic protein Bcl-2 showed decreasing trend (F=0.324,P<0.05).
Conclusion　The ent-kaurane diterpenoid J32 can inhibit the migration and proliferation of pancreatic cancer SW1990 cells in a concentration dependent manner,promote apoptosis and increase the level of intracellular reactive oxygen species;in addition,J32 can promote DNA damage and cell cycle arrest of cells,and its mechanism may be related to the ATR-Chk1-p53-Cyclin D1 pathway.

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更新日期/Last Update: 2023-09-05