Role of trefoil factor3/phosphatidylinositl 3-kinase/protein kinase B/nuclear factorκB in the proliferation and apoptosis of human papillary thyroid carcinoma cell line TPC-1

GUO Meng-Yao; SUN Lin; LIN Xu; ZHANG Wen-Jing; ZHANG Jing; WU Jing-Fang; XUE Gang

doi:10.16098/j.issn.0529-1356.2020.04.009

PDF(2832 KB)

Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (4) : 528-535. DOI: 10.16098/j.issn.0529-1356.2020.04.009

Role of trefoil factor3/phosphatidylinositl 3-kinase/protein kinase B/nuclear factorκB in the proliferation and apoptosis of human papillary thyroid carcinoma cell line TPC-1

GUO Meng-yao¹ SUN Lin¹ LIN Xu¹ZHANG Wen-jing¹ZHANG Jing¹ WU Jing-fang^1* XUE Gang^2*

Author information +

History +

Abstract

Objective To investigate the effects of trefoil factor 3 (TFF3) on the proliferation and apoptosis of human thyroid papillary carcinoma cell line (TPC-1) and its molecular mechanism. Methods The lentiviral expression vector of overexpression and knockdown of TFF3 gene were constructed，293T cell was packaged to produce lentiviral particles, virus solution was collected and transfected into TPC-1 cells, enhanced cell TFF3-TPC-1 and enhanced control group ConTFF3-TPC-1; silencing cell shRNA-TFF3-TPC-1 and silencing control cells shCon -TPC-1. Western blotting, and Real-time PCR were used to detect the expression of TFF3 protein and mRNA of four groups. Growth curve and colony formation assay were used to detect the proliferation.Flow cytometry was used to analyze the apoptosis level of the four groups; Western blotting and immunocytochemistry were used to detect apoptosis-related protein and pathway protein phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), nuclear factor-κB（NF-κB）expression. Results 1. Overexpression and inhibition of expression of TFF3 stable cell TFF3-TPC-1 and shRNA-TFF3-TPC-1 were constructed suscessfully. 2. The proliferation and cloning ability of TFF3-TPC-1 cells were significantly higher than those of ConTFF3-TPC-1 cells（P＜0.05 or P＜0.01）, the proliferation and cloning ability of shRNA-TFF3-TPC-1 cells were significantly lower than those of shCon-TPC-1 cells(P<0.01); 3. The apoptosis rate of TFF3-TPC-1 cells was lower than that of ConTFF3-TPC-1 (0.75%±0.08% vs 5.62%±0.3%, P<0.01),and the apoptosis rate of shRNA-TFF3-TPC-1 was higher than that of shConTPC-1 (22.2% ± 1.2% vs 5.34% ± 0.4%, P<0.01); 4. After silencing TFF3 gene, the expressions of Bax, cytochrome C (Cyt-C), cleavedCaspase-9, cleaved-Caspase-3 were up-regulated, and the expressions of Bcl-2, Akt, p-Akt and NF-κB-P65 were downregulated (P<0.05 or P<0.01). Conclusion TFF3 may regulate the proliferation and apoptosis of TPC-1 cells by affecting the PI3K/Akt/NF-κB signaling pathway.

Key words

Trefoil factor 3 / phosphatidylinositol 3-kinase/protein kinase B/nuclear factor kappa B / Papillary thyroid carcinoma / Lentiviral transfection / Western blotting / Flow cytometry / Real-time PCR / Human

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

GUO Meng-yao SUN Lin LIN Xu ZHANG Wen-jing ZHANG Jing WU Jing-fang XUE Gang. Role of trefoil factor3/phosphatidylinositl 3-kinase/protein kinase B/nuclear factorκB in the proliferation and apoptosis of human papillary thyroid carcinoma cell line TPC-1[J]. Acta Anatomica Sinica. 2020, 51(4): 528-535 https://doi.org/10.16098/j.issn.0529-1356.2020.04.009

References

［1］ Fitzmaurice C, Akinyemiju TF, Al Lami FH, et al.Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 29 cancer groups, 1990 to 2016:a systematic analysis for the Global Burden of Disease Study［J］. JAMA Oncol, 2018，4(11):1553-1568.

［2］ Liu J, Kim SY, Shin S,et al.Overexpression of TFF3 is involved in prostate carcinogenesis via blocking mitochondria-mediatedapoptosis［J］. Exp Mol Med, 2018, 50(8):104-111.

［3］ Yuan Z, Chen D, Chen X, et al. Overexpression of trefoil factor 3 (TFF3) contributes to the malignant progression in cervical cancer cells［J］. Cancer Cell Int, 2017, 17:7-19.

［4］ Gro?e-Kreul J, Busch M, Winter C, et al.Forced trefoil factor family peptide 3 (TFF3) expression reduces growth, viability, and tumorigenicity of human retinoblastoma cell lines［J］. PLoS One, 2016,11(9):e0163025.

［5］ Dai J, Lin X, Guo MY, et al.Effect and molecular mechanism of trefoil factor family 3 on the cell cycle of thyroid papillary carcinoma cell［J］.Acta Anatomica Sinica,2019,50(2):211-219. (in Chinese)

代金,林旭,郭梦姚,等.三叶因子3对甲状腺乳头状癌细胞周期的影响及机制［J］.解剖学报,2019,50(2):211-219.

［6］ Lin X,Zhang H,Dai J et al.TFF3 Contributes to epithelial-mesenchymal transition (EMT) in papillary thyroid carcinoma cells via the MAPK/ERK signaling pathway［J］. J Cancer,2018,9(23): 4430-4439.

［7］ Wu J, Zhang H, Zhang W, et al. Lentivirus-mediated shRNA interference of trefoil factor 3 blocks cell viability, migration and invasion in the papillary thyroid carcinoma cells.［J］. Neoplasma, 2018, 65(2):169.

［8］ Hanisch C, Sharbati J, Kutz-Lohroff B, et al. Trefoil factor 3 mediates resistance to apoptosis in colon carcinoma cells by a regulatory RNA axis［J］. Cell Death and Disease, 2017, 8(3):e2660.

［9］ Cheng J. Modern Apoptosis Molecular Biology［M］. 3rd Edition. Beijing: Science Press, 2017:36-45.(in Chinese)

成军. 现代细胞凋亡分子生物学［M］.第3版. 北京：科学出版社, 2017:36-45.

［10］ You ML, Chen YJ, Chong QY, et al. Trefoil factor 3 mediation of oncogenicity and chemoresistance in hepatocellular carcinoma is AKT-BCL-2 dependent［J］. Oncotarget, 2017, 8(24):39323-39344.

［11］ Wu LP, Xu LY, HU XF，et al Effect of cod skin oligopeptide on the proliferation and apoptosis in human gastric cancer cells line SGC-7901 and its mechanisml［J］.Acta Anatomica Sinica,2017,48(6):698-703.(in Chinese)

吴丽萍,续力云,胡晓斐,等.鳕鱼皮寡肽对人胃癌细胞SGC-7901增殖凋亡的影响及机制［J］.解剖学报,2017,48(6):698-703.

［12］ Yin J, Ji Z, Hong Y, et al. ShMARCH8 Inhibits tumorigenesis via PI3K pathway in gastric cancer［J］.Cell Physiol Biochem, 2018, 49(1):306-321.

［13］ Chen C, Liu TS, Zhao SC,et al.XIAP impairs mitochondrial function during apoptosis by regulating the Bcl-2 family in renal cell carcinoma［J］. Exp Ther Med, 2018,15(5): 4587-4593.

［14］ Wang N, Hou MS, Zhan Y, et al. MALAT1 promotes cisplatin resistance in cervical cancer by activating the PI3K/AKT pathway［J］.Eur Rev Med Pharmacol Sci, 2018,22(22):7653-7659.

［15］ Zhou H, Xu J, Wang S,et al. Role of cantharidin in the activation of IKKα/ⅠκBα/NFκB pathway by inhibiting PP2A activity in cholangiocarcinoma cell lines［J］. Mol Med Rep, 2018,17(6):7672-7682.

［16］ Han Z, Hong L, Han Y, et al. Phospho Akt mediates multidrug resistance of gastric cancer cells through regulation of P-gp, Bcl-2 and Bax［J］. J Exp Clin Cancer Res, 2007, 26(2):261-268.

［17］ Zhou B, Wang D, Sun G, et al Effect of miR-21 on apoptosis in lung cancer cell through Inhibiting the PI3K/Akt/NF-κB signaling pathway in vitro and in vivo［J］. Cell Physiol Biochem, 2018, 46(3):999-1008.

［18］ Chen H, Huang Y, Huang J, et al.Gigantol attenuates the proliferation of human liver cancer HepG2 cellsthrough the PI3K/Akt/NF-κB signaling pathway［J］. Oncol Rep, 2017, 37(2):865-870.

［19］ Han B, Cui PH, Zhao GJ, et al. Effects of silencing S6K2 on proliferation, apoptosis and PI3K/Akt/NF-κB pathway in cervical carcinoma cells［J］.Chinese Clinical Oncology, 2018, 23(10):28-34.(in Chinese)

韩冰, 崔彭华, 赵国军,等.S6K2沉默对宫颈癌细胞增殖凋亡及PI3K/Akt/NF-κB通路的影响［J］. 临床肿瘤学杂志, 2018, 23(10):28-34.

［20］ Huang, WS,Huang, CY,Hsieh, MC, et al.Expression of PRDX6 correlates with migration and invasiveness of colorectal cancer cells［J］. Cell Physiol Biochem, 2018,51(6):2616-2630.