FAM3C通过激活Akt促进口腔鳞癌细胞活力
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摘要
目的:探讨FAM3C (family with sequence similarity 3, member C)蛋白在口腔鳞癌细胞中的表达和作用。方法:通过RT-qPCR及Western blot法检测人口腔鳞癌癌前病变细胞DOK和口腔鳞癌细胞WSU-HN6中FAM3C的mRNA及蛋白表达水平。在WSU-HN6细胞中分别使用siFAM3C和抗FAM3C抗体处理,在DOK细胞中使用腺病毒过表达FAM3C,分别在处理后24、48和72 h使用CCK-8和Western blot法检测FAM3C对细胞活力及蛋白激酶B(protein kinase B,Akt)激活的影响。结果:(1)与DOK细胞相比,WSU-HN6细胞中FAM3C的mRNA和蛋白表达水平显著升高(P<0.05);(2)siFAM3C在转染后48 h和72 h均可抑制WSU-HN6细胞的活力(P<0.05),同时抑制Akt的激活(P<0.05);(3)抗体封闭FAM3C蛋白48 h和72 h也能抑制WSU-HN6细胞的活力,并抑制Akt的激活(P<0.05);(4)过表达FAM3C 48 h和72 h可促进DOK细胞的活力,并激活Akt(P<0.05)。结论:FAM3C可能通过激活Akt促进口腔鳞癌细胞的活力。
AIM: To investigate the expression and roles of family with sequence similarity 3,member C( FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte( DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with si FAM3C or FAM3C antibody. After 24,48 and 72 h,the viability of WSU-HN6 cells was measured by CCK-8 assay,and the activation of protein kinase B( Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24,48 and 72 h,and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells,the mRNA and protein levels of FAM3C were significantly increased in the WSUHN6 cells( P < 0. 05). The viability of WSU-HN6 cells transfected with si FAM3C was significantly inhibited at 48 h and72 h( P < 0. 05). si FAM3C treatment inhibited the activation of Akt( P < 0. 05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt( P < 0. 05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt( P < 0. 05). CONCLUSION:FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.
AIM: To investigate the expression and roles of family with sequence similarity 3,member C( FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte( DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with si FAM3C or FAM3C antibody. After 24,48 and 72 h,the viability of WSU-HN6 cells was measured by CCK-8 assay,and the activation of protein kinase B( Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24,48 and 72 h,and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells,the mRNA and protein levels of FAM3C were significantly increased in the WSUHN6 cells( P < 0. 05). The viability of WSU-HN6 cells transfected with si FAM3C was significantly inhibited at 48 h and72 h( P < 0. 05). si FAM3C treatment inhibited the activation of Akt( P < 0. 05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt( P < 0. 05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt( P < 0. 05). CONCLUSION:FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.
引文
[1] Bavle RM, Venugopal R, Konda P, et al. Molecular classification of oral squamous cell carcinoma[J]. J Clin Diagn Res, 2016, 10(9):ZE18-ZE21.
[2] Thomson PJ. Oral carcinogenesis[M]// Thomson PJ. Oral precancer: diagnosis and management of potentially malignant disorders. 1st ed. Chichester: Wiley-Blackwell, 2012:31-47.
[3] Hasegawa H, Liu L, Tooyama I, et al. The FAM3 superfamily member ILEI ameliorates Alzheimer's disease-like pathology by destabilizing the penultimate amyloid-beta precursor[J]. Nat Commun, 2014, 5:3917.
[4] Gao ZH, Lu C, Wang ZN, et al. ILEI: a novel marker for epithelial-mesenchymal transition and poor prognosis in colorectal cancer[J]. Histopathology, 2014, 65(4):527-538.
[5] Gronborg M, Kristiansen TZ, Iwahori A, et al. Biomarker discovery from pancreatic cancer secretome using a diffe-rential proteomic approach[J]. Mol Cell Proteomics, 2006, 5(1):157-171.
[6] Zhu YH, Zhang B, Li M, et al. Prognostic significance of FAM3C in esophageal squamous cell carcinoma[J]. Diagn Pathol, 2015,10:192.
[7] Wu CC, Xiao Y, Li H, et al. Overexpression of FAM3C is associated with poor prognosis in oral squamous cell carcinoma[J]. Pathol Res Pract, 2019 Jan 14. [Epub adead of print]
[8] Franke TF. PI3K/Akt: getting it right matters[J]. Oncogene, 2008, 27(50):6473-6488.
[9] Chen Z, Ding L, Yang W, et al. Hepatic activation of the FAM3C-HSF1-CaM pathway attenuates hyperglycemia of obese diabetic mice[J]. Diabetes, 2017, 66(5):1185-1197.
[10] Chen Z, Wang J, Yang W, et al. FAM3C activates HSF1 to suppress hepatic gluconeogenesis and attenuate hyperglycemia of type 1 diabetic mice[J]. Oncotarget, 2017, 8(62):106038-106049.
[11] Waerner T, Alacakaptan M, Tamir I, et al. ILEI: a cytokine essential for EMT, tumor formation, and late events in metastasis in epithelial cells[J]. Cancer Cell, 2006, 10(3):227-239.
[12] Lahsnig C, Mikula M, Petz M, et al. ILEI requires oncogenic Ras for the epithelial to mesenchymal transition of hepatocytes and liver carcinoma progression[J]. Oncogene, 2009, 28(5):638-650.
[13] Shi M, Duan G, Nie S, et al. Elevated FAM3C promotes cell epithelial-mesenchymal transition and cell migration in gastric cancer[J]. Onco Targets Ther, 2018, 11:8491-8505.
[14] Zhang X, Feng H, Li Z, et al. Aspirin is involved in the cell cycle arrest, apoptosis, cell migration, and invasion of oral squamous cell carcinoma[J]. Int J Mol Sci, 2018, 19(7):E2029.
[15] Csiszar A, Kutay B, Wirth S, et al. Interleukin-like epithelial-to-mesenchymal transition inducer activity is controlled by proteolytic processing and plasminogen-urokinase plasminogen activator receptor system-regulated secretion during breast cancer progression[J]. Breast Cancer Res, 2014, 16:433.
[16] Janda E, Litos G, Grunert S, et al. Oncogenic Ras/Her-2 mediate hyperproliferation of polarized epithelial cells in 3D cultures and rapid tumor growth via the PI3K pathway[J]. Oncogene, 2002, 21(33):5148-5159.
[17] Janda E, Lehmann K, Killisch I, et al. Ras and TGFβ cooperatively regulate epithelial cell plasticity and metastasis: dissection of Ras signaling pathways[J]. J Cell Biol, 2002, 156(2):299-313.
[18] Kral M, Klimek C, Kutay B, et al. Covalent dimerization of interleukin-like epithelial-to-mesenchymal transition (EMT) inducer (ILEI) facilitates EMT, invasion, and late aspects of metastasis[J]. FEBS J, 2017, 284(20):3484-3505.
[19] Noguchi K, Dalton AC, Howley BV, et al. Interleukin-like EMT inducer regulates partial phenotype switching in MITF-low melanoma cell lines[J]. PLoS One, 2017, 12(5): e0177830.
[20] Li Y, Wang J, Wang F, et al. Tissue microarray analysis reveals the expression and prognostic significance of phosphorylated AktThr308 in oral squamous cell carcinoma[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2013, 116(5):591-597.
[21] Matsuo FS, Andrade MF, Loyola AM, et al. Pathologic significance of AKT, mTOR, and GSK3β proteins in oral squamous cell carcinoma-affected patients[J]. Virchows Arch, 2018, 472(6):983-997.
[22] Wang Y, Lin L, Xu H, et al. Genetic variants in AKT1 gene were associated with risk and survival of OSCC in Chinese Han population[J]. J Oral Pathol Med, 2015, 44(1):45-50.
[23] Chang F, Lee JT, Navolanic PM, et al. Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy[J]. Leukemia, 2003, 17(3):590-603.
[24] Zhao X, Luo G, Fan Y, et al. ILEI is an important intermediate participating in the formation of TGF-β1-induced renal tubular EMT[J]. Cell Biochem Funct, 2018, 36(2):46-55.
[2] Thomson PJ. Oral carcinogenesis[M]// Thomson PJ. Oral precancer: diagnosis and management of potentially malignant disorders. 1st ed. Chichester: Wiley-Blackwell, 2012:31-47.
[3] Hasegawa H, Liu L, Tooyama I, et al. The FAM3 superfamily member ILEI ameliorates Alzheimer's disease-like pathology by destabilizing the penultimate amyloid-beta precursor[J]. Nat Commun, 2014, 5:3917.
[4] Gao ZH, Lu C, Wang ZN, et al. ILEI: a novel marker for epithelial-mesenchymal transition and poor prognosis in colorectal cancer[J]. Histopathology, 2014, 65(4):527-538.
[5] Gronborg M, Kristiansen TZ, Iwahori A, et al. Biomarker discovery from pancreatic cancer secretome using a diffe-rential proteomic approach[J]. Mol Cell Proteomics, 2006, 5(1):157-171.
[6] Zhu YH, Zhang B, Li M, et al. Prognostic significance of FAM3C in esophageal squamous cell carcinoma[J]. Diagn Pathol, 2015,10:192.
[7] Wu CC, Xiao Y, Li H, et al. Overexpression of FAM3C is associated with poor prognosis in oral squamous cell carcinoma[J]. Pathol Res Pract, 2019 Jan 14. [Epub adead of print]
[8] Franke TF. PI3K/Akt: getting it right matters[J]. Oncogene, 2008, 27(50):6473-6488.
[9] Chen Z, Ding L, Yang W, et al. Hepatic activation of the FAM3C-HSF1-CaM pathway attenuates hyperglycemia of obese diabetic mice[J]. Diabetes, 2017, 66(5):1185-1197.
[10] Chen Z, Wang J, Yang W, et al. FAM3C activates HSF1 to suppress hepatic gluconeogenesis and attenuate hyperglycemia of type 1 diabetic mice[J]. Oncotarget, 2017, 8(62):106038-106049.
[11] Waerner T, Alacakaptan M, Tamir I, et al. ILEI: a cytokine essential for EMT, tumor formation, and late events in metastasis in epithelial cells[J]. Cancer Cell, 2006, 10(3):227-239.
[12] Lahsnig C, Mikula M, Petz M, et al. ILEI requires oncogenic Ras for the epithelial to mesenchymal transition of hepatocytes and liver carcinoma progression[J]. Oncogene, 2009, 28(5):638-650.
[13] Shi M, Duan G, Nie S, et al. Elevated FAM3C promotes cell epithelial-mesenchymal transition and cell migration in gastric cancer[J]. Onco Targets Ther, 2018, 11:8491-8505.
[14] Zhang X, Feng H, Li Z, et al. Aspirin is involved in the cell cycle arrest, apoptosis, cell migration, and invasion of oral squamous cell carcinoma[J]. Int J Mol Sci, 2018, 19(7):E2029.
[15] Csiszar A, Kutay B, Wirth S, et al. Interleukin-like epithelial-to-mesenchymal transition inducer activity is controlled by proteolytic processing and plasminogen-urokinase plasminogen activator receptor system-regulated secretion during breast cancer progression[J]. Breast Cancer Res, 2014, 16:433.
[16] Janda E, Litos G, Grunert S, et al. Oncogenic Ras/Her-2 mediate hyperproliferation of polarized epithelial cells in 3D cultures and rapid tumor growth via the PI3K pathway[J]. Oncogene, 2002, 21(33):5148-5159.
[17] Janda E, Lehmann K, Killisch I, et al. Ras and TGFβ cooperatively regulate epithelial cell plasticity and metastasis: dissection of Ras signaling pathways[J]. J Cell Biol, 2002, 156(2):299-313.
[18] Kral M, Klimek C, Kutay B, et al. Covalent dimerization of interleukin-like epithelial-to-mesenchymal transition (EMT) inducer (ILEI) facilitates EMT, invasion, and late aspects of metastasis[J]. FEBS J, 2017, 284(20):3484-3505.
[19] Noguchi K, Dalton AC, Howley BV, et al. Interleukin-like EMT inducer regulates partial phenotype switching in MITF-low melanoma cell lines[J]. PLoS One, 2017, 12(5): e0177830.
[20] Li Y, Wang J, Wang F, et al. Tissue microarray analysis reveals the expression and prognostic significance of phosphorylated AktThr308 in oral squamous cell carcinoma[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2013, 116(5):591-597.
[21] Matsuo FS, Andrade MF, Loyola AM, et al. Pathologic significance of AKT, mTOR, and GSK3β proteins in oral squamous cell carcinoma-affected patients[J]. Virchows Arch, 2018, 472(6):983-997.
[22] Wang Y, Lin L, Xu H, et al. Genetic variants in AKT1 gene were associated with risk and survival of OSCC in Chinese Han population[J]. J Oral Pathol Med, 2015, 44(1):45-50.
[23] Chang F, Lee JT, Navolanic PM, et al. Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy[J]. Leukemia, 2003, 17(3):590-603.
[24] Zhao X, Luo G, Fan Y, et al. ILEI is an important intermediate participating in the formation of TGF-β1-induced renal tubular EMT[J]. Cell Biochem Funct, 2018, 36(2):46-55.