紫苏醇对结肠癌SW480细胞凋亡的影响及其机制研究
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摘要
目的研究紫苏醇对结肠癌SW480细胞凋亡的影响及其可能机制。方法体外培养结肠癌SW480细胞,分为不同浓度(1μg·mL~(-1)、5μg·mL~(-1)、10μg·mL~(-1))阿帕替尼组和不同浓度(1μg·mL~(-1)、5μg·mL~(-1)、10μg·mL~(-1))紫苏醇组,检测各组细胞内Rspo3和Notch-1蛋白的表达、细胞增殖活性、细胞周期、细胞侵袭、转移能力以及凋亡蛋白Caspase-3的活性。结果紫苏醇和阿帕替尼均可抑制结肠癌SW480细胞内Rspo3和Notch-1蛋白的表达,并随着浓度升高,抑制作用呈增强趋势(P<0.05),且紫苏醇对Rspo3和Notch-1蛋白表达的抑制作用显著优于相同剂量的阿帕替尼(P<0.05)。紫苏醇和阿帕替尼均可抑制结肠癌SW480细胞的增殖和侵袭,且抑制作用随浓度升高而增强(P<0.05)。紫苏醇和阿帕替尼处理后,G_0/G_1期细胞显著增加(P<0.05),而G_2/M期细胞无显著变化(P>0.05)。低剂量(1μg·mL~(-1))紫苏醇和阿帕替尼对Caspase 3活性无明显影响(P>0.05),而中、高剂量(5μg·mL~(-1)、10μg·mL~(-1))紫苏醇和阿帕替尼可显著增强结肠癌SW480细胞中Caspase-3的活性(P<0.05)。结论紫苏醇可抑制结肠癌SW480细胞的增殖和细胞内Notch-1蛋白的表达,并反馈性下调Rspo3基因与蛋白的表达。
Objective To study the effect of perillyl alcohol on apoptosis of colon cancer SW480 cells, and to reveal the specific mechanism of perillyl alcohol-induced apoptosis through Rspo3 and Notch signaling pathways. Methods Colon cancer SW480 cells were treated with different concentrations of apatinib(1, 5, 10 μg·mL~(-1)) or perillyl alcohol(1, 5, 10 μg·mL~(-1)). Intracellular Rspo3 and Notch-1 expressions were detected by Western-blot method. Detect the cell proliferation activity, cycle, invasion and metastasis ability, as well as the activity of apoptosis protein Caspase-3. Results Both perillyl alcohol and apatinib inhibited the expression of intracellular Rspo3 and Notch-1 and enhanced their inhibitory ability with increasing concentration(P<0.05). The inhibiting effect of perillyl alcohol on protein expression of Rspo3 and Notch-1 was much greater than that of apatinib at the same dose(P<0.05). Both perillyl alcohol and apatinib inhibited the proliferation and invasion of colon cancer SW480 cells,and their inhibitory ability was strengthened with the increase of concentrations(P<0.05). G_0/G_1 phase cells were significantly increased after treatment with perillyl alcohol and apatinib(P<0.05), but there was no significant change in G_2/M phase cells(P>0.05). Perillyl alcohol and apafittin could not inhibit Caspase 3 activity at low dose(1 μg·mL~(-1))(P>0.05), but significantly increased the Caspase-3 activity in colon cancer SW480 cells at medium and high doses(5, 10 μg·mL~(-1))(P<0.05). Conclusion Perillyl alcohol can inhibit the proliferation of colon cancer SW480 cells and the expression of intracellular Notch-1 protein, and down-regulate the expression of Rspo3 gene and protein through feedback.
Objective To study the effect of perillyl alcohol on apoptosis of colon cancer SW480 cells, and to reveal the specific mechanism of perillyl alcohol-induced apoptosis through Rspo3 and Notch signaling pathways. Methods Colon cancer SW480 cells were treated with different concentrations of apatinib(1, 5, 10 μg·mL~(-1)) or perillyl alcohol(1, 5, 10 μg·mL~(-1)). Intracellular Rspo3 and Notch-1 expressions were detected by Western-blot method. Detect the cell proliferation activity, cycle, invasion and metastasis ability, as well as the activity of apoptosis protein Caspase-3. Results Both perillyl alcohol and apatinib inhibited the expression of intracellular Rspo3 and Notch-1 and enhanced their inhibitory ability with increasing concentration(P<0.05). The inhibiting effect of perillyl alcohol on protein expression of Rspo3 and Notch-1 was much greater than that of apatinib at the same dose(P<0.05). Both perillyl alcohol and apatinib inhibited the proliferation and invasion of colon cancer SW480 cells,and their inhibitory ability was strengthened with the increase of concentrations(P<0.05). G_0/G_1 phase cells were significantly increased after treatment with perillyl alcohol and apatinib(P<0.05), but there was no significant change in G_2/M phase cells(P>0.05). Perillyl alcohol and apafittin could not inhibit Caspase 3 activity at low dose(1 μg·mL~(-1))(P>0.05), but significantly increased the Caspase-3 activity in colon cancer SW480 cells at medium and high doses(5, 10 μg·mL~(-1))(P<0.05). Conclusion Perillyl alcohol can inhibit the proliferation of colon cancer SW480 cells and the expression of intracellular Notch-1 protein, and down-regulate the expression of Rspo3 gene and protein through feedback.
引文
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[12]Zhao J, Lin W, Cao Z, et al. Total alkaloids of Rubus alceifolius Poir inhibit tumor angiogenesis through suppression of the Notch signaling pathway in a mouse model of hepatocellular carcinoma[J]. Mol Med Rep, 2015, 11(1):357-361.doi:10.3892/mmr.2014.2702.
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[14]倪晓辰,赵志红,马永良,等.桧木醇诱导人肾透明细胞癌786-O细胞凋亡及其机制研究[J].中国肿瘤临床, 2015, 42(1):43-46. doi:10.3969/j.issn.1000-8179.20141412.
[15]王晶.秦皮甲素对人肺癌细胞A549凋亡影响[J].中国公共卫生, 2015, 31(4):464-466. doi:10.11847/zgggws2015-31-04-25.
[2]王敏,崔博豪,孙舒雅,等.四种蛋白类肿瘤标志物对结直肠癌筛查的价值[J].世界华人消化杂志, 2017, 25(11):996-1001. doi:10.11569/wcjd. v25. i11.996.
[3]郑金琪,王前,杨希,等.药物代谢酶及其亚型介导的抗肿瘤药物临床毒性反应研究进展[J].中国药学杂志,2017, 52(13):1115-1119. doi:10.11669/cpj.2017.13.004.
[4]周玭,李筠,刘云章,等.一个新的Wnt信号通路相关蛋白——Rspo3[J].国际病理科学与临床杂志, 2011, 31(4):344-349. doi:10.3969/j. issn.1673-2588.2011.04.015.
[5]Dormoy V, Jacqmin D, Lang H, et al. From development to cancer:lessons from the kidney to uncover new therapeutic targets[J]. Anticancer Res, 2012, 32(9):3609-3617. doi:10.1016/j. pdpdt.2012.07.004.
[6]Gong X, Yi J, Carmon KS, et al. Aberrant RSPO3-LGR4signaling in Keap1-deficient lung adenocarcinomas promotes tumor aggressiveness[J]. Oncogene, 2015, 34(36):4692-4701. doi:10.1038/onc.2014.417.
[7]刘艳华,王小中.外泌体在疾病诊疗中的研究进展[J].实验与检验医学, 2014, 32(3):267-271. doi:10.3969/j.issn.1674-1129.2014.03.012.
[8]周录平.腹腔镜结肠癌根治术的临床疗效[J].中国老年学, 2013, 33(21):5327-5328. doi:10.3969/j. issn.1005-9202.2013.21.052.
[9]Hao HX, Jiang X, Cong F. Control of Wnt Receptor Turnover by R-spondin-ZNRF3/RNF43 Signaling Module and Its Dysregulation in Cancer[J]. Cancers(Basel), 2016, 8(6).pii:E54. doi:10.3390/cancers8060054.
[10]Han T, Schatoff EM, Murphy C, et al. R-Spondin chromosome rearrangements drive Wnt-dependent tumour initiation and maintenance in the intestine[J]. Nat Commun,2017, 8:15945. doi:10.1038/ncomms15945.
[11]林琳,周刘祥,王晓春.MiRNA-424对人结肠癌细胞系Lovo细胞增殖及Rspo3基因表达的影响[J].中国肿瘤, 2018, 27(3):223-228. doi:10.11735/j.issn.1004-0242.2018.03. A012.
[12]Zhao J, Lin W, Cao Z, et al. Total alkaloids of Rubus alceifolius Poir inhibit tumor angiogenesis through suppression of the Notch signaling pathway in a mouse model of hepatocellular carcinoma[J]. Mol Med Rep, 2015, 11(1):357-361.doi:10.3892/mmr.2014.2702.
[13]Arora A, Bansal A. The importance of microvessel density in predicting cancer progression in patients with penile squamous cell carcinoma[J]. Int Urol Nephrol, 2017, 49(6):1007-1014. doi:10.1007/s11255-017-1565-3.
[14]倪晓辰,赵志红,马永良,等.桧木醇诱导人肾透明细胞癌786-O细胞凋亡及其机制研究[J].中国肿瘤临床, 2015, 42(1):43-46. doi:10.3969/j.issn.1000-8179.20141412.
[15]王晶.秦皮甲素对人肺癌细胞A549凋亡影响[J].中国公共卫生, 2015, 31(4):464-466. doi:10.11847/zgggws2015-31-04-25.