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Effect of FAM83D silencing on radiosensitivity of esophageal squamous cell carcinoma cells by mediating epithelial-mesenchymaltransition
Yang Xingxiao1, Zou Naiyi2, Zhang Xueyuan2, Ma Ming3, Zhu Shuchai2
1Department of Infection Management, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China; 2Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China; 3Department of Laboratory, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China
AbstractObjective To examine the effect of FAM83D knockdown on proliferation, survival ability and invasion of human esophageal squamous cell carcinoma after X-ray radiation, and explore the mechanism. Methods The expression of FAM83D, E-cadherin and vimentin in tumor tissues was detected in 69 cases of esophageal squamous cell cancer by using immunohistochemical method. The siRNA based on the sequences of the FAM83D mRNA were synthesized to transfect into the cultured ECA109 cells as FAM83D shRNA group. The effect of silencing FAM83D gene was evaluated to determine the protein levels of FAM83D in the human oesophageal squamous cell carcinoma ECA109 and KYSE30 cells using western blotting. MTS, clone formation, and Transwell assay were employed to examine the proliferation, survival ability and invasion of ECA109 and KYSE30 cells in vitro, respectively. We used flow cytometry assay to analyze distribution of cell apoptosis in different groups. Western blotting was used to examine the expression of cell metastasis-related molecules and apoptosis-related protein. Results The strong expression rates of FAM83D, E-cadherin, and vimentin were 55%(38/69), 36%(25/69) and 61%(42/69) in the tumor tissues, respectively. FAM83D protein expression was significantly and negatively correlated with the expression of E-cadherin (r=-0.350, P<0.01), and positively with the expression of vimentin (r=0.470,P<0.01). Western blotting results demonstrated that silencing FAM83D gene significantly reduced the FAM83D protein expression (P<0.01). MTS data demonstrated that FAM83D knockdown after irradiation significantly inhibited the proliferation of esophageal squamous cell carcinoma ECA109 and KYSE30 cells (P<0.05). The data from the clone formation assay revealed that the radiosensitivity was increased after downragulation of FAM83D expression (P<0.01). In addition, the invasive abilities of oesophageal carcinoma cells transfected with FAM83D shRNA after irradiation were significantly inhibited compared with those of the NC group (P<0.01), followed by the downregulation of N-cadherin, vimentin, Snail, p-Akt and p-GSK-3β expression, and the upregulation of E-cadherin expression (P<0.01). The apoptosis rate of tumor cells in FAM83D shRNA group after irradiation was markedly increased (P<0.01), followed by a decrease of Bcl-2 and Mcl-1 expression and an increase of Cleaved caspase-3 expression (P<0.01). Conclusions FAM83D expressions was found to be closely related to the invasion and development of ESCC. Furthermore, siRNA interference technology inhibited the expression of FAM83D gene in oesophageal squamous cell carcinoma cells, reduced the proliferation, invasion of cells, induced cell apoptosis, and increased radiosensitivity, which may be associated with regulating the epithelial-mesenchymaltransition via Snail/Akt/GSK-3β signaling pathways.
Fund:National Natural Science Foundation of China (81872456, 81903118);National Natural Science Foundation of Hebei (H2020206292);Hebei Institute of Medical Sciences (20180483)
Yang Xingxiao,Zou Naiyi,Zhang Xueyuan et al. Effect of FAM83D silencing on radiosensitivity of esophageal squamous cell carcinoma cells by mediating epithelial-mesenchymaltransition[J]. Chinese Journal of Radiation Oncology, 2021, 30(10): 1071-1077.
Yang Xingxiao,Zou Naiyi,Zhang Xueyuan et al. Effect of FAM83D silencing on radiosensitivity of esophageal squamous cell carcinoma cells by mediating epithelial-mesenchymaltransition[J]. Chinese Journal of Radiation Oncology, 2021, 30(10): 1071-1077.
[1] Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015[J]. CA Cancer J Clin, 2016, 66(2):115-132. DOI:10.3322/caac.21338. [2] Siegel RL, Fedewa SA, Miller KD, et al. Cancer statistics for Hispanics/Latinos, 2015[J]. CA Cancer J Clin, 2015, 65(6):457-480. DOI:10.3322/caac.21314. [3] Stark TW, Hensley PJ, Spear A, et al. Predictive value of epithelial-mesenchymal-transition (EMT) signature and PARP-1 in prostate cancer radioresistance[J]. Prostate, 2017, 77(16):1583-1591. DOI:10.1002/pros.23435. [4] Zhang Q, Yu S, Lok SIS, et al. FAM83D promotes ovarian cancer progression and its potential application in diagnosis of invasive ovarian cancer[J]. J Cell Mol Med, 2019, 23(7):4569-4581. DOI:10.1111/jcmm.14360. [5] Shi R, Sun J, Sun Q, et al. Upregulation of FAM83D promotes malignant phenotypes of lung adenocarcinoma by regulating cell cycle[J]. Am J Cancer Res, 2016, 6(11):2587-2598. [6] Wang D, Han S, Peng R, et al. FAM83D activates the MEK/ERK signaling pathway and promotes cell proliferation in hepatocellular carcinoma[J]. Biochem Biophys Res Commun, 2015, 458(2):313-320. DOI:10.1016/j.bbrc.2015.01.108. [7] Wang Z, Liu Y, Zhang P, et al. FAM83D promotes cell proliferation and motility by downregulating tumor suppressor gene FBXW7[J]. Oncotarget, 2013, 4(12):2476-2486. DOI:10.18632/oncotarget.1581. [8] Yin C, Lin X, Wang Y, et al. FAM83D promotes epithelial-mesenchymal transition, invasion and cisplatin resistance through regulating the AKT/mTOR pathway in non-small-cell lung cancer[J]. Cell Oncol (Dordr), 2020, 43(3):395-407. DOI:10.1007/s13402-020-00494-9. [9] Liu Y, Chen LH, Yuan YW, et al. Activation of AKT is associated with metastasis of nasopharyngeal carcinoma[J]. Tumour Biol, 2012, 33(1):241-245. DOI:10.1007/s13277-011-0272-4. [10] Dong C, Wu Y, Yao J, et al. G9a interacts with Snail and is critical for Snail-mediated E-cadherin repression in human breast cancer[J]. J Clin Invest, 2012, 122(4):1469-1486. DOI:10.1172/JCI57349. [11] Zhu H, Diao S, Lim V, et al. FAM83D inhibits autophagy and promotes proliferation and invasion of ovarian cancer cells via PI3K/AKT/mTOR pathway[J]. Acta Biochim Biophys Sin (shanghai), 2019, 51(5):509-516. DOI:10.1093/abbs/gmz028. [12] Mu Y, Zou H, Chen B, et al. FAM83D knockdown regulates proliferation, migration and invasion of colorectal cancer through inhibiting FBXW7/Notch-1 signalling pathway[J]. Biomed Pharmacother, 2017, 90:548-554. DOI:10.1016/j.biopha.2017.03.073. [13] Karami H, Baradaran B, Esfehani A, et al. Down-regulation of Mcl-1 by small interference RNA induces apoptosis and sensitizes HL-60 leukemia cells to etoposide[J]. Asian Pac J Cancer Prev, 2014, 15(2):629-635. DOI:10.7314/ apjcp.2014.15.2.629. [14] Katayama S, Shimoda K, Takenaga Y. Loss of ADAR1 in human iPS cells promotes caspase-3-mediated apoptotic cell death[J]. Genes Cells, 2015, 20(8):675-680. DOI:10.1111/gtc.12261.
2021, Vol. 30 
