下调解耦联蛋白2表达抑制Fadu细胞增殖
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摘要
目的探讨解耦联蛋白2(UCP2)对下咽癌Fadu细胞的影响,及UCP2对细胞周期以及细胞增殖的影响;同时探讨临床常用化疗药物顺铂(cisplatin)对UCP2基因下调后的Fadu细胞增殖的影响。方法流式细胞术/PI染色法检测UCP2基因对Fadu细胞周期的影响;集落形成实验检测UCP2基因下调对Fadu细胞增殖的影响;MTT法和Brdu法检测UCP2能否增强Fadu细胞对顺铂的敏感性以及使用UCP2抑制剂京平尼(genipin)处理细胞后,Fadu细胞的增殖情况。结果干扰UCP2表达抑制Fadu细胞周期于G_1/S期;抑制细胞集落形成,但干扰UCP2表达对cisplatin抑制Fadu增殖无协同作用;genipin抑制UCP2的表达后,细胞增殖受到抑制,具有时间/浓度依赖性。结论下调UCP2的表达后,Fadu细胞的增殖能力减弱,且抑制细胞周期于G_1/S期,提示UCP2可能在下咽癌Fadu细胞增殖过程中发挥重要的调节作用。
Objective The effect of uncoupling protein 2(UCP2) on the Fadu cells of the hypopharyngeal cancer was investigated in this study. The siRNA technology was used to study the effect of UCP2 on cell cycle and cell proliferation. Meanwhile, cisplatin, a commonly chemotherapeutic drug, was researched on the effect of proliferation in Fadu cells after down-regulated UCP2 gene. Methods Flow cytometry/PI staining was used to observe the effect of UCP2 on Fadu cell cycle. Clone formation assay was used to detect the effect of down-regulated UCP2 on the proliferation of Fadu cells. MTT assay and Brud assay were used to determine whether UCP2 enhanced the sensitivity of Fadu cells to cisplatin and to detect the effect of UCP2 on the proliferation of Fadu. Results The present result showed that down-regulated UCP2 expression inhibited Fadu cell cycle in G_1/S phase and cell colony formation, but had no synergistic effect on inhibition of Fadu proliferation by cisplatin; The expression of UCP2 was significantly inhibited by genipin, and then inhibited cell proliferation in a time-concentration-dependent manner. Conclusion The proliferation of Fadu cells was weakened and the cell cycle was inhibited in G_1/S phase after down-regulated UCP2, suggesting that UCP2 may play an important adjust role in the development in Fadu of hypopharyngeal cancer.
Objective The effect of uncoupling protein 2(UCP2) on the Fadu cells of the hypopharyngeal cancer was investigated in this study. The siRNA technology was used to study the effect of UCP2 on cell cycle and cell proliferation. Meanwhile, cisplatin, a commonly chemotherapeutic drug, was researched on the effect of proliferation in Fadu cells after down-regulated UCP2 gene. Methods Flow cytometry/PI staining was used to observe the effect of UCP2 on Fadu cell cycle. Clone formation assay was used to detect the effect of down-regulated UCP2 on the proliferation of Fadu cells. MTT assay and Brud assay were used to determine whether UCP2 enhanced the sensitivity of Fadu cells to cisplatin and to detect the effect of UCP2 on the proliferation of Fadu. Results The present result showed that down-regulated UCP2 expression inhibited Fadu cell cycle in G_1/S phase and cell colony formation, but had no synergistic effect on inhibition of Fadu proliferation by cisplatin; The expression of UCP2 was significantly inhibited by genipin, and then inhibited cell proliferation in a time-concentration-dependent manner. Conclusion The proliferation of Fadu cells was weakened and the cell cycle was inhibited in G_1/S phase after down-regulated UCP2, suggesting that UCP2 may play an important adjust role in the development in Fadu of hypopharyngeal cancer.
引文
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[21] Imai K,Fukuda T,Wada T,et al.UCP2 expression may represent a predictive marker of neoadjuvant chemotherapy effectiveness for locally advanced uterine cervical cancer[J].Oncol Lett,2017,14(1):951-957.
[22] Wang RH,Yung KC,Zhao YJ,et al.A mechanism for the temporal potentiation of genipin to the cytotoxicity of cisplatin in colon cancer cells[J].Int J Med Sci,2016,13(7):507-516.
[23] Li Z,Zhang TB,Jia DH,et al.Genipin inhibits the growth of human bladder cancer cellsvia inactivation of PI3K/Akt signaling[J].Oncol Lett,2018,15(2):2619-2624.
[24] Bo RK,Jeong YA,Na YJ,et al.Genipin suppresses colorectal cancer cells by inhibiting the Sonic Hedgehog pathway[J].Oncotarget,2017,8 (60):101952-101964.
[25] Kim ES,Jeong CS,Moon A.Genipin,a constituent of Gardenia jasminoides Ellis,induces apoptosis and inhibits invasion in MDA-MB-231 breast cancer cells[J].Oncol Rep,2012,27(2):567-572.
[2] Argiris A,Kamlnouzis MV,Raben D,et al.Head and neck cancer[J].Lancet,2008,371(9625):1695-1709.
[3] Hanna GJ,Haddad RI,Lorch JH.Induction chemotherapy for locoregionally advanced head and neck cancer:past,present,future[J]?Oncologist,2013,18(3):288-293.
[4] Pitt MA.Overexpression of uncoupling protein-2 in cancer:metabolic and heat changes,inhibition and effects on drug resistance[J].Inflammopharmacology,2015,23(6):365-369.
[5] Li W,Nichols K,Nathan CA,et al.Mitochondrial uncoupling protein 2 is up-regulated in human head and neck,skin,pancreatic,and prostate tumors[J].Cancer Biomark,2013,13(5):377-383.
[6] Wang MX,Li GY,Yang ZW,et al.Uncoupling protein 2 downregulation by hypoxia through repression of peroxisome proliferator-activated receptor g promotes chemoresistance of non-small cell lung cancer[J].Oncotarget,2017,8(5):8083-8094.
[7] Cruz MD,Ledbetter S,Chowdhury S,et al.Metabolic reprogramming of the premalignant colonic mucosa is an early event in carcinogenesis[J].Oncotarget,2017,8(13):20543-20557.
[8] Pitt MA.Overexpression of uncoupling protein-2 in cancer:metabolic and heat changes,inhibition and effects on drug resistance[J].Inflammopharmacology,2015,23(6):365-369.
[9] Li W,Zhang C,Jackson K,et al.UCP2 Knockout Suppresses Mouse Skin Carcinogenesis[J].Cancer Prev Res (Phila),2015,8(6):487-491.
[10] Zhou L.Pay attention to the standardized treatment of laryngeal cancer and hypopharyngeal cancer[J].Chinese Journal of Otorhinolaryngology Head and Neck Surgery,2009,44(9):705-706.(in Chinese)周梁.重视喉癌和下咽癌的规范化治疗[J].中华耳鼻咽喉头颈外科杂志,2009,44(9):705-706.
[11] Wang Y,Zhang HY,Ying GG,et al.Expression of UCP2 in gastric adenocarcinoma tissues and its effect on biological function of gastric adenocarcinoma cells[J].Cancer Res Prev Treat,2018,45(4):215-220.(in Chinese)王艺,张海洋,应国光,等.胃腺癌组织中UCP2的表达及其对癌细胞生物学功能的影响[J].肿瘤防治研究,2018,45(4):215-220.
[12] An JD,Jiang Y,Bai YF,et al.Effect of uncoupling protein 2 on activation of hepatic stellate cell in liver fibrosis of rats[J].Acta Anatomica Sinica,2014,45(4):480-484.(in Chinese)安建多,江瑛,白云飞,等.解耦联蛋白2对大鼠肝纤维化形成中星形细胞活化的影响[J].解剖学报,2014,45(4):480-484.
[13] Toda C,Diano S.Mitochondrial UCP2 in the central regulation of metabolism[J].Best Pract Res Clin Endocrinol Metab,2014,28(5):757-764.
[14] Lou Y,Liu S,Zhang C,et al.Enhanced atherosclerosis in TIPE2-deficient mice is associated with increased macrophage responses to oxidized low-density lipoprotein[J].J Immunol,2013,191(9):4849-4857.
[15] Yamamoto K,Lee BJ,Li C,et al.Early B-cell-specific inactivation of ATM synergizes with ectopic CyclinD1 expression to promote pre-germinal center B-cell lymphomas in mice[J].Leukemia,2015,29(6):1414-1424.
[16] Liu W,Yin TF,Ren JH,et al.Activation of the EGFR/Akt/NF-KB/cyclinD1 survival signaling pathway in human cholesteatoma epithelium[J].Eur Arch Otorhinolaryngol,2014,1(2):265-273.
[17] Liang S,Mu K,Wang Y,et al.CyclinD1,a prominent prognostic marker for endometrial diseases[J].Diagn Pathol,2013,8:138.
[18] Zhuang MJ,Liu D,Chen YW,et al.Astragalus polysaccharide inhibiting Lewis lung carcinoma transplanted tumor in mice and influences the expression of Caspase-3 and Smac /Diablo[J].Acta Anatomica Sinica,2018,49(1):63-69.(in Chinese)庄梦婕,刘丹,陈彦文,等.黄芪多糖联合顺铂对小鼠 Lewis 肺癌移植Caspase-3、Smac / Diablo 表达的影响[J].解剖学报,2018,49(1):63-69.
[19] Amable L.Cisplatin resistance and opportunities for precision medicine[J].Pharmacol Res,2016,106:27-36.
[20] Esteves P,Pecqueur C,Ransy C,et al.Mitochondrial retrograde signaling mediated by UCP2 inhibits cancer cell proliferation and tumorigenesis[J].Cancer Res,2014,74(14):3971-3982.
[21] Imai K,Fukuda T,Wada T,et al.UCP2 expression may represent a predictive marker of neoadjuvant chemotherapy effectiveness for locally advanced uterine cervical cancer[J].Oncol Lett,2017,14(1):951-957.
[22] Wang RH,Yung KC,Zhao YJ,et al.A mechanism for the temporal potentiation of genipin to the cytotoxicity of cisplatin in colon cancer cells[J].Int J Med Sci,2016,13(7):507-516.
[23] Li Z,Zhang TB,Jia DH,et al.Genipin inhibits the growth of human bladder cancer cellsvia inactivation of PI3K/Akt signaling[J].Oncol Lett,2018,15(2):2619-2624.
[24] Bo RK,Jeong YA,Na YJ,et al.Genipin suppresses colorectal cancer cells by inhibiting the Sonic Hedgehog pathway[J].Oncotarget,2017,8 (60):101952-101964.
[25] Kim ES,Jeong CS,Moon A.Genipin,a constituent of Gardenia jasminoides Ellis,induces apoptosis and inhibits invasion in MDA-MB-231 breast cancer cells[J].Oncol Rep,2012,27(2):567-572.