Knockdown of ZFPL1 results in increased autophagy and autophagy‑related cell death in NCI‑N87 and BGC‑823 human gastric carcinoma cell lines

Xie,Yong‑Zheng; Ma,Wan‑Li; Meng,Ji‑Ming; Ren,Xue‑Qun

doi:10.3892/mmr.2017.6300

Knockdown of ZFPL1 results in increased autophagy and autophagy‑related cell death in NCI‑N87 and BGC‑823 human gastric carcinoma cell lines

Authors:
- Yong‑Zheng Xie
- Wan‑Li Ma
- Ji‑Ming Meng
- Xue‑Qun Ren
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Affiliations: Department of General Surgery, Huaihe Hospital of Henan University, Kaifeng, Henan 475000, P.R. China
Published online on: March 9, 2017 https://doi.org/10.3892/mmr.2017.6300
Pages: 2633-2642

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Abstract

Macroautophagy, which will hereafter be referred to as autophagy, is an evolutionarily conserved process, during which cells recycle and remove damaged organelles and proteins in response to cellular stress. However, the mechanisms underlying the regulation of autophagy remain to be fully elucidated. The present study demonstrated that knockdown of zinc finger protein like 1 (ZFPL1) induces autophagy and increases autophagic cell death in NCI‑N87 and BGC‑823 human gastric carcinoma cell lines. To examine the role of ZFPL1 in gastric carcinoma cells, ZFPL1 expression was downregulated by lentiviral infection. Zinc finger domain‑FLAG was used to compete with ZFPL1 for golgin A2/GM130 binding. Autophagy was analyzed by red fluorescent protein‑microtubule‑associated protein 1A/1B‑light chain 3 (LC3) puncta, LC3I to LC3II conversion, and p62 expression. The results demonstrated that knockdown of ZFPL1 was able to significantly increase cell death rate. However, ZFPL1 knockdown exerted almost no effect on the expression of apoptosis‑associated markers, including B cell lymphoma 2 (Bcl‑2), Bcl‑x, Bcl‑2‑associated X protein, BH3 interacting domain death agonist, p53, and the classical caspase family members, caspase‑3, caspase‑8 and caspase‑9. An endogenous ZFPL1‑GM130 association was identified in NCI‑N87 cells and BGC‑823 cells by co‑immunoprecipitation. Furthermore, cell death was restricted following treatment of ZFPL1 knockdown cells with an autophagy inhibitor. Therefore, knockdown of ZFPL1 expression may induce cell death via autophagy, rather than apoptosis. These results suggest that ZFPL1 may serve an important role in regulating autophagy in NCI‑N87 and BGC‑823 cells.

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1	Tan P and Yeoh KG: Genetics and molecular pathogenesis of gastric adenocarcinoma. Gastroenterology. 149:1153–1162.e3. 2015. View Article : Google Scholar : PubMed/NCBI
2	Wesolowski R, Lee C and Kim R: Is there a role for second-line chemotherapy in advanced gastric cancer? Lancet Oncol. 10:903–912. 2009. View Article : Google Scholar : PubMed/NCBI
3	Takahashi T, Saikawa Y and Kitagawa Y: Gastric cancer: Current status of diagnosis and treatment. Cancers. 5:48–63. 2013. View Article : Google Scholar : PubMed/NCBI
4	Bhutia SK, Das SK, Azab B, Dash R, Su ZZ, Lee SG, Dent P, Curiel DT, Sarkar D and Fisher PB: Autophagy switches to apoptosis in prostate cancer cells infected with melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24). Autophagy. 7:1076–1077. 2011. View Article : Google Scholar : PubMed/NCBI
5	Janku F, McConkey DJ, Hong DS and Kurzrock R: Autophagy as a target for anticancer therapy. Nat Rev Clin Oncol. 8:528–539. 2011. View Article : Google Scholar : PubMed/NCBI
6	Klionsky DJ and Emr SD: Autophagy as a regulated pathway of cellular degradation. Science. 290:1717–1721. 2000. View Article : Google Scholar : PubMed/NCBI
7	Levine B and Kroemer G: Autophagy in the pathogenesis of disease. Cell. 132:27–42. 2008. View Article : Google Scholar : PubMed/NCBI
8	Klionsky DJ, Codogno P, Cuervo AM, Deretic V, Elazar Z, Fueyo-Margareto J, Gewirtz DA, Kroemer G, Levine B, Mizushima N, et al: A comprehensive glossary of autophagy-related molecules and processes. Autophagy. 6:438–448. 2010. View Article : Google Scholar : PubMed/NCBI
9	Zhang J, Li Y, Chen X, Liu T, Chen Y, He W, Zhang Q and Liu S: Autophagy is involved in anticancer effects of matrine on SGC-7901 human gastric cancer cells. Oncol Rep. 26:115–124. 2011.PubMed/NCBI
10	Zhang QY, Wu LQ, Zhang T, Han YF and Lin X: Autophagy-mediated HMGB1 release promotes gastric cancer cell survival via RAGE activation of extracellular signal-regulated kinases 1/2. Oncol Rep. 33:1630–1638. 2015.PubMed/NCBI
11	Thedieck K, Holzwarth B, Prentzell MT, Boehlke C, Kläsener K, Ruf S, Sonntag AG, Maerz L, Grellscheid SN, Kremmer E, et al: Inhibition of mTORC1 by astrin and stress granules prevents apoptosis in cancer cells. Cell. 154:859–874. 2013. View Article : Google Scholar : PubMed/NCBI
12	Sun Y, Liu JH, Jin L, Sui YX, Han LL and Huang Y: Effect of autophagy-related beclin1 on sensitivity of cisplatin-resistant ovarian cancer cells to chemotherapeutic agents. Asian Pac J Cancer Prev. 16:2785–2791. 2015. View Article : Google Scholar : PubMed/NCBI
13	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
14	Chiu CF, Ghanekar Y, Frost L, Diao A, Morrison D, McKenzie E and Lowe M: ZFPL1, a novel ring finger protein required for cis-Golgi integrity and efficient ER-to-Golgi transport. EMBO J. 27:934–947. 2008. View Article : Google Scholar : PubMed/NCBI
15	Nakamura N, Rabouille C, Watson R, Nilsson T, Hui N, Slusarewicz P, Kreis TE and Warren G: Characterization of a cis-Golgi matrix protein, GM130. J Cell Biol. 131:1715–1726. 1995. View Article : Google Scholar : PubMed/NCBI
16	Keith SA, Maddux SK, Zhong Y, Chinchankar MN, Ferguson AA, Ghazi A and Fisher AL: Graded proteasome dysfunction in Caenorhabditis elegans activates an adaptive response involving the conserved SKN-1 and ELT-2 transcription factors and the autophagy-lysosome pathway. PLoS Genet. 12:e10058232016. View Article : Google Scholar : PubMed/NCBI
17	Chang SH, Hong SH, Jiang HL, Minai-Tehrani A, Yu KN, Lee JH, Kim JE, Shin JY, Kang B, Park S, et al: GOLGA2/GM130, cis-Golgi matrix protein, is a novel target of anticancer gene therapy. Mol Ther. 20:2052–2063. 2012. View Article : Google Scholar : PubMed/NCBI
18	Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, Agholme L, Agnello M, Agostinis P, Aguirre-Ghiso JA, et al: Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 8:445–544. 2012. View Article : Google Scholar : PubMed/NCBI
19	Levine B: Cell biology: Autophagy and cancer. Nature. 446:745–747. 2007. View Article : Google Scholar : PubMed/NCBI
20	Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, Øvervatn A, Bjørkøy G and Johansen T: p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem. 282:24131–24145. 2007. View Article : Google Scholar : PubMed/NCBI
21	Kepp O, Galluzzi L, Lipinski M, Yuan J and Kroemer G: Cell death assays for drug discovery. Nat Rev Drug Discov. 10:221–237. 2011. View Article : Google Scholar : PubMed/NCBI
22	Wang J, Whiteman MW, Lian H, Wang G, Singh A, Huang D and Denmark T: A non-canonical MEK/ERK signaling pathway regulates autophagy via regulating Beclin 1. J Biol Chem. 284:21412–21424. 2009. View Article : Google Scholar : PubMed/NCBI
23	Wu YC, Wu WK, Li Y, Yu L, Li ZJ, Wong CC, Li HT, Sung JJ and Cho CH: Inhibition of macroautophagy by bafilomycin A1 lowers proliferation and induces apoptosis in colon cancer cells. Biochem Biophys Res Commun. 382:451–456. 2009. View Article : Google Scholar : PubMed/NCBI
24	Fulda S: Targeting extrinsic apoptosis in cancer: Challenges and opportunities. Semin Cell Dev Biol. 39:20–25. 2015. View Article : Google Scholar : PubMed/NCBI
25	Reed JC: Apoptosis-targeted therapies for cancer. Cancer Cell. 3:17–22. 2003. View Article : Google Scholar : PubMed/NCBI
26	Galluzzi L, Pedro Bravo-San JM, Vitale I, Aaronson SA, Abrams JM, Adam D, Alnemri ES, Altucci L, Andrews D, Annicchiarico-Petruzzelli M, et al: Essential versus accessory aspects of cell death: Recommendations of the NCCD 2015. Cell Death Differ. 22:58–73. 2015. View Article : Google Scholar : PubMed/NCBI
27	Gewirtz DA: Cytoprotective and nonprotective autophagy in cancer therapy. Autophagy. 9:1263–1265. 2013. View Article : Google Scholar : PubMed/NCBI
28	Gewirtz DA: The four faces of autophagy: Implications for cancer therapy. Cancer Res. 74:647–651. 2014. View Article : Google Scholar : PubMed/NCBI
29	Fulda S and Kögel D: Cell death by autophagy: Emerging molecular mechanisms and implications for cancer therapy. Oncogene. 34:5105–5113. 2015. View Article : Google Scholar : PubMed/NCBI
30	Li Z, Wang J and Yang X: Functions of autophagy in pathological cardiac hypertrophy. Int J Biol Sci. 11:672–678. 2015. View Article : Google Scholar : PubMed/NCBI
31	Galluzzi L, Pietrocola F, Levine B and Kroemer G: Metabolic control of autophagy. Cell. 159:1263–1276. 2014. View Article : Google Scholar : PubMed/NCBI
32	Pupyshev AB: Reparative autophagy and autophagy death of cells. Functional and regulatory aspects. Tsitologiia. 56:179–196. 2014.(In Russian). PubMed/NCBI
33	Pinho SS, Carvalho S, Marcos-Pinto R, Magalhães A, Oliveira C, Gu J, Dinis-Ribeiro M, Carneiro F, Seruca R and Reis CA: Gastric cancer: Adding glycosylation to the equation. Trends Mol Med. 19:664–676. 2013. View Article : Google Scholar : PubMed/NCBI
34	Zavareh Beheshti R, Lau KS, Hurren R, Datti A, Ashline DJ, Gronda M, Cheung P, Simpson CD, Liu W, Wasylishen AR, et al: Inhibition of the sodium/potassium ATPase impairs N-glycan expression and function. Cancer Res. 68:6688–6697. 2008. View Article : Google Scholar : PubMed/NCBI
35	Contessa JN, Bhojani MS, Freeze HH, Ross BD, Rehemtulla A and Lawrence TS: Molecular imaging of N-linked glycosylation suggests glycan biosynthesis is a novel target for cancer therapy. Clin Cancer Res. 16:3205–3214. 2010. View Article : Google Scholar : PubMed/NCBI
36	Vitale I, Manic G, Dandrea V and De Maria R: Role of autophagy in the maintenance and function of cancer stem cells. Int J Dev Biol. 59:95–108. 2015. View Article : Google Scholar : PubMed/NCBI
37	Hoppener JW, De Wit MJ, Simarro-Doorten AY, Roijers JF, van Herrewaarden HM, Lips CJ, Parente F, Quincey D, Gaudray P, Khodaei S, et al: A putative human zinc-finger gene (ZFPL1) on 11q13, highly conserved in the mouse and expressed in exocrine pancreas. The European Consortium on MEN 1. Genomics. 50:251–259. 1998. View Article : Google Scholar : PubMed/NCBI