薏苡仁油诱导胃癌细胞SGC-7901凋亡的实验研究
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摘要
目的探讨薏苡仁活性成分诱导胃癌细胞(SGC-7901)凋亡的可能机制。方法将不同浓度的薏苡仁油(2、4、8 mg/mL)作用于SGC-7901细胞,采用MTT法检测药物对细胞增殖的影响,流式细胞术检测药物诱导细胞的凋亡,划痕实验检测薏苡仁油抑制细胞的迁移,Transwell小室检测药物抑制细胞的侵袭,Western blot检测相关蛋白PRMT5、PI3K和AKT的表达。结果 MTT结果显示,2、4 mg/mL的薏苡仁油能显著抑制SGC-7901细胞的增殖,2 mg/mL薏苡仁油抑制率达(30. 02±1. 56)%,与空白对照组相比差异极显著(P <0. 01)。流式细胞实验结果表明,2、4 mg/mL的薏苡仁油细胞凋亡率分别为(16. 25±2. 54)%、(12. 60±1. 12)%,与空白对照组(2. 00±1. 22)%相比差异极显著(P <0. 01)。细胞划痕实验结果表明,2、4 mg/mL的薏苡仁油处理过的SGC-7901细胞迁移缓慢,与空白对照组间距对比差异极显著(P <0. 01)。侵袭实验结果表明,2、4 mg/mL的薏苡仁油可显著抑制细胞的侵袭,细胞侵袭数分别是(134. 00±2. 86)、(167. 00±0. 99)个,与对照组的(268. 00±2. 05)个相比,差异极显著(P <0. 01),8 mg/mL组的迁移数为(203. 00±2. 97)个,与对照组对比差异显著(P <0. 05)。Western blot结果显示,不同浓度的薏苡仁油能够显著下调SGC-7901细胞中PRMT5、PI3K和AKT的表达。结论薏苡仁油能够显著抑制胃癌SGC-7901细胞的增殖、迁移与侵袭,其机制可能是通过下调PRMT5-PI3K/AKT信号通路,阻断下游多种抗凋亡分子的活化,诱导细胞凋亡、抑制胃癌SGC-7901细胞的侵袭与转移。
Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells( SGC-7901). Methods Different concentrations of coix seed oil( 2,4,8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation,and flow cytometry for drug-induced cell apoptosis,scratch test for cell migration inhibited by coix seed oil,Transwell chamber for drug-inhibited cell invasion,and Western blot for expression of related proteins PRMT5,PI3 K and AKT. Results The results of MTT showed that 2,4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells,and the inhibition rate of 2 mg/mL was( 30. 02 ± 1. 56) %,which showed significant difference compared to the control group( P < 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were( 16. 25 ± 2. 54) %,( 12. 60 ± 1. 12) %,respectively,and which showed a significant difference compared with( 2. 0 ± 1. 22) % in the control group( P < 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2,4 mg/mL of coix seed oil had significant difference when compared to control group( P <0. 01). The results of invasion experiments showed that 2,4 mg/mL of coix seed oil could significantly inhibit cell invasion,and the number of cell invasion was( 134. 00 ± 2. 86),( 167. 00 ±0. 99),respectively,which showed significant difference compared to( 268. 00 ± 2. 05) in the control group( P < 0. 01). The migration number in 8 mg/mL coix seed oil group was( 167 ± 0. 99),a significant difference was observed when compared to the control group( P < 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5,PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation,migration and invasion of gastric cancer SGC-7901 cells,its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules,induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.
Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells( SGC-7901). Methods Different concentrations of coix seed oil( 2,4,8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation,and flow cytometry for drug-induced cell apoptosis,scratch test for cell migration inhibited by coix seed oil,Transwell chamber for drug-inhibited cell invasion,and Western blot for expression of related proteins PRMT5,PI3 K and AKT. Results The results of MTT showed that 2,4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells,and the inhibition rate of 2 mg/mL was( 30. 02 ± 1. 56) %,which showed significant difference compared to the control group( P < 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were( 16. 25 ± 2. 54) %,( 12. 60 ± 1. 12) %,respectively,and which showed a significant difference compared with( 2. 0 ± 1. 22) % in the control group( P < 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2,4 mg/mL of coix seed oil had significant difference when compared to control group( P <0. 01). The results of invasion experiments showed that 2,4 mg/mL of coix seed oil could significantly inhibit cell invasion,and the number of cell invasion was( 134. 00 ± 2. 86),( 167. 00 ±0. 99),respectively,which showed significant difference compared to( 268. 00 ± 2. 05) in the control group( P < 0. 01). The migration number in 8 mg/mL coix seed oil group was( 167 ± 0. 99),a significant difference was observed when compared to the control group( P < 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5,PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation,migration and invasion of gastric cancer SGC-7901 cells,its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules,induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.
引文
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[16]黄晖婷,李劲高,肖定璋,等.手霉素通过阻断PI3K/Akt信号通路诱导U937白血病细胞凋亡[J].实用医学杂志,2017,33(7):1066-1069.
[17] L V X Y,M A L,CHEN J F,et al. Knockdown of DUXAP10 inhibits proliferation and promotes apoptosis in bladder cancer cells via PI3K/Akt/m TOR signaling pathway[J]. Int J Oncol,2018,52(1):288-294.
[18] LIU J,CHEN W,ZHANG H,et al. miR-214 targets the PTEN-mediated PI3K/Akt signaling pathway and regulates cell proliferation and apoptosis in ovarian cancer[J]. Oncol Lett,2017,14(5):5711-5718.
[19]孔桂美,王继军,陶文华,等.组蛋白精氨酸甲基转移酶5在胃癌组织中的表达及其意义[J].吉林大学学报:医学版,2016,42(4):753-756.
[20] Zhan Y P,Huang X E,Cao J,et al. Clinical safety and efficacy of kanglaite(coix seed oil)injection combined with chemotherapy in treating patients with gastric cancer[J]. Asian Pacific Journal of Cancer Prevention,2012,13(10):5319-5321.
[21]朴商均.论中药的双向调节[D].济南:山东中医药大学,2003.
[22]边晶,李洲.双向调节作用理论与临床研究进展[J].中国城乡企业卫生,2016,31(9):49-51.
[23]徐志立,于彩娜,张明波,等.中药对“脾胃运化水液”功能双向调节的分子机制及其研究模式的建立与应用[J].辽宁中医杂志,2018,45(5):944-948.
[2] Okada E,Ukawa S,Nakamura K,et al. Demographic and lifestyle factors and survival among patients with esophageal and gastric cancer:The Biobank Japan Project[J]. Journal of Epidemiology,2017,27(3):S29-S35.
[3] Chen W Q,Zheng R S,Baade P D,et al. Cancer statistics in China,2015[J]. CA:A Cancer Journal for Clinicians,2016,66(2):115-132.
[4] Sada Y H,Smaglo B G,Tran Cao H S,et al. National trends in multimodality therapy for locally advanced gastric cancer[J]. Journal of Surgical Research,2019,237:41-49.
[5] Okada K,Oka Y,Kawanaka S,et al. Three cases of advanced gastric cancer with peritoneal dissemination successfully treated with S-1 and docetaxel combination chemotherapy[J]. Gan To Kagaku Ryoho,2018,45(13):2252-2254.
[6] Quan P,Zheng P Y,You S F,et al. Clinical and psychometric validation of the quality of life assessment system for advanced gastric cancer based on traditional Chinese medicine[J]. Chin J Integr Med,2016,22(8):581-588.
[7] Zhang P R,Meng X Y,Tang X H,et al. The effect of a coix seed oil injection on cancer pain relief[J]. Support Care Cancer,2019,27(2):461-465.
[8]房玮,吴红卫,李莹莹,等.中药联合化疗治疗胃癌的临床效果分析[J].实用癌症杂志,2015,30(11):1668-1671.
[9] Liu X,Xiu L J,Jiao J P,et al. Traditional Chinese medicine integrated with chemotherapy for stage IV non-surgical gastric cancer:a retrospective clinical analysis[J]. Journal of Integrative Medicine,2017,15(6):469-475.
[10]梁欣妍,丁筑红.薏苡仁油对消化系统肿瘤的药理作用及临床应用[J].实用医学杂志,2017,33(1):159-161.
[11]苏芳,于常华.薏苡仁油抗恶性肿瘤作用的研究进展[J].临床荟萃,2014,29(9):1075-1078.
[12] Liu Y,Zhang W,Wang X J,et al. Antitumor effect of Kanglaite injection in human pancreatic cancer xenografts[J].BMC Complement Altern Med,2014,14:228-234.
[13] Wang D,Yang C D,Wang Z E,et al. Norcantharidin combined with Coix seed oil synergistically induces apoptosis and inhibits hepatocellular carcinoma growth by downregulating regulatory T cells accumulation[J]. Sci Rep,2017,7(1):9373-9385.
[14] Bedford M T,Richard S. Arginine methylation[J]. Molecular Cell,2005,18(3):263-272.
[15] Wei T Y W,Juan C C,Hisa J Y,et al. Protein arginine methyltransferase 5 is a potential oncoprotein that upregulates G1 cyclins/cyclin-dependent kinases and the phosphoinositide3-kinase/AKT signaling cascade[J]. Cancer Sci,2012,103(9):1640-1650.
[16]黄晖婷,李劲高,肖定璋,等.手霉素通过阻断PI3K/Akt信号通路诱导U937白血病细胞凋亡[J].实用医学杂志,2017,33(7):1066-1069.
[17] L V X Y,M A L,CHEN J F,et al. Knockdown of DUXAP10 inhibits proliferation and promotes apoptosis in bladder cancer cells via PI3K/Akt/m TOR signaling pathway[J]. Int J Oncol,2018,52(1):288-294.
[18] LIU J,CHEN W,ZHANG H,et al. miR-214 targets the PTEN-mediated PI3K/Akt signaling pathway and regulates cell proliferation and apoptosis in ovarian cancer[J]. Oncol Lett,2017,14(5):5711-5718.
[19]孔桂美,王继军,陶文华,等.组蛋白精氨酸甲基转移酶5在胃癌组织中的表达及其意义[J].吉林大学学报:医学版,2016,42(4):753-756.
[20] Zhan Y P,Huang X E,Cao J,et al. Clinical safety and efficacy of kanglaite(coix seed oil)injection combined with chemotherapy in treating patients with gastric cancer[J]. Asian Pacific Journal of Cancer Prevention,2012,13(10):5319-5321.
[21]朴商均.论中药的双向调节[D].济南:山东中医药大学,2003.
[22]边晶,李洲.双向调节作用理论与临床研究进展[J].中国城乡企业卫生,2016,31(9):49-51.
[23]徐志立,于彩娜,张明波,等.中药对“脾胃运化水液”功能双向调节的分子机制及其研究模式的建立与应用[J].辽宁中医杂志,2018,45(5):944-948.