Knockdown of FAM225B inhibits the progression of the hypertrophic scar following glaucoma surgery by inhibiting autophagy

Ma,Xianpeng; Liu,Lili

doi:10.3892/mmr.2021.11843

Knockdown of FAM225B inhibits the progression of the hypertrophic scar following glaucoma surgery by inhibiting autophagy

Authors:
- Xianpeng Ma
- Lili Liu
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Affiliations: Department of Dermatology, Affiliated Hospital of Beihua University, Jilin, Jilin 132001, P.R. China
Published online on: January 14, 2021 https://doi.org/10.3892/mmr.2021.11843
Article Number: 204
Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The formation of a hypertrophic scar (HS) may lead to failure of glaucoma surgery. Long non‑coding RNAs (lncRNAs) are involved in the formation of HSs. Moreover, family with sequence similarity 225 member B (FAM225B) is upregulated in HS. However, the role of the lncRNA FAM225B in HS remains unknown. Thus, the present study aimed to investigate the function of FAM225B in HS. Scar fibroblasts were isolated from patients who had undergone glaucoma surgery. Western blotting was used to detect the expressions of Bax, Bcl‑2, cleaved caspase 3, p62, ATG7 and Beclin 1, and reverse transcription‑quantitative PCR (RT‑qPCR) were conducted to determine the level of FAM225B in scar fibroblasts. Microtubule associated protein 1 light chain 3 α staining was performed to examine autophagosomes in scar fibroblasts. Furthermore, cell proliferation was evaluated via 5‑ethynyl‑2'‑deoxyuridine staining. Flow cytometry was conducted to determine cell apoptosis and the levels of reactive oxygen species (ROS) in scar fibroblasts. The cell migratory ability was assessed using a Transwell assay. The results demonstrated that FAM225B knockdown significantly attenuated scar fibroblast proliferation and induced apoptosis. Additionally, transfection of scar fibroblasts with FAM225B small interfering RNA (siRNA) significantly increased the ROS levels and significantly decreased the migration of scar fibroblasts. The FAM225B overexpression‑induced increase of scar fibroblast proliferation and migration was significantly reversed by 3‑methyladenine administration. The results suggested that knockdown of FAM225B significantly inhibited the proliferation of scar fibroblasts by inhibiting autophagy. Therefore, knockdown of FAM225B could inhibit scar fibroblast proliferation after glaucoma surgery by inhibiting autophagy. These findings may provide a novel perspective of developing treatment strategy for the patients with HSs after glaucoma surgery.

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1	Li X, Leng Y, Jiang Q, Wang Z, Luo P, Zhang C, Chen L, Wang Y, Wang H, Yue X, et al: Eye drops of metformin prevents fibrosis after glaucoma filtration surgery in rats via activating AMPK/Nrf2 signaling pathway. Front Pharmacol. 11:10382020. View Article : Google Scholar : PubMed/NCBI
2	Swogger J, Conner IP, Rosano M, Kemmerer M, Happ-Smith C, Wells A, Schuman JS and Yates CC: Injected versus sponge-applied mitomycin C (MMC) during modified trabeculectomy in New Zealand white rabbit model. Transl Vis Sci Technol. 9:232020. View Article : Google Scholar : PubMed/NCBI
3	Yaman D, Takmaz T, Yüksel N, Dincer SA and Şahin FI: Evaluation of silent information regulator T (SIRT) 1 and Forkhead Box O (FOXO) transcription factor 1 and 3a genes in glaucoma. Mol Biol Rep. 47:9337–9344. 2020. View Article : Google Scholar : PubMed/NCBI
4	Cheng WS, Chen CL, Chen JT, Lin LT, Pao SI, Chen YH and Lu DW: AR12286 alleviates TGF-β-related myofibroblast transdifferentiation and reduces fibrosis after glaucoma filtration surgery. Molecules. 25:44222020. View Article : Google Scholar
5	Yamamoto K, Kokubun T, Sato K, Akaishi T, Shimazaki A, Nakamura M, Shiga Y, Tsuda S, Omodaka K, Saya H and Nakazawa T: The DNA topoisomerase II inhibitor amsacrine as a novel candidate adjuvant in a model of glaucoma filtration surgery. Sci Rep. 9:192882019. View Article : Google Scholar : PubMed/NCBI
6	Yu J, Qiu LX, Qing GP, Zhao BW and Wang H: Modified cortex mori capsules improving the successful rate of functional filtering blebs after reclinical glaucoma filtering surgery. World J Clin Cases. 7:3436–3445. 2019. View Article : Google Scholar : PubMed/NCBI
7	Zhang F, Liu K, Pan Z, Cao M, Zhou D, Liu H, Huang Y and Duan X: Effects of rosiglitazone/PHBV drug delivery system on postoperative fibrosis in rabbit glaucoma filtration surgery model. Drug Deliv. 26:812–819. 2019. View Article : Google Scholar : PubMed/NCBI
8	Sun J, Liu X, Lei Y, Tang M, Dai Z, Yang X, Yu X, Yu L, Sun X and Ding J: Sustained subconjunctival delivery of cyclosporine A using thermogelling polymers for glaucoma filtration surgery. J Mater Chem B. 5:6400–6411. 2017. View Article : Google Scholar : PubMed/NCBI
9	Holló G: Wound Healing and Glaucoma Surgery: Modulating the scarring process with conventional antimetabolites and new molecules. Dev Ophthalmol. 59:80–89. 2017. View Article : Google Scholar : PubMed/NCBI
10	Sharma A, Anumanthan G, Reyes M, Chen H, Brubaker JW, Siddiqui S, Gupta S, Rieger FG and Mohan RR: Epigenetic modification prevents excessive wound healing and scar formation after glaucoma filtration surgery. Invest Ophthalmol Vis Sci. 57:3381–3389. 2016. View Article : Google Scholar : PubMed/NCBI
11	Zhang H, Li J, Shao W and Shen N: lncRNA SNHG9 is downregulated in osteoarthritis and inhibits chondrocyte apoptosis by downregulating miR-34a through methylation. BMC Musculoskelet Disord. 21:5112020. View Article : Google Scholar : PubMed/NCBI
12	Wang JH, Wu XJ, Duan YZ and Li F: Circular RNA_CNST promotes the tumorigenesis of osteosarcoma cells by sponging miR-421. Cell Transplant. 29:9636897209261472020. View Article : Google Scholar : PubMed/NCBI
13	Xu YT, Leng YR, Liu MM, Dong RF, Bian J, Yuan LL, Zhang JG, Xia YZ and Kong LY: MicroRNA and long noncoding RNA involvement in gout and prospects for treatment. Int Immunopharmacol. 87:1068422020. View Article : Google Scholar : PubMed/NCBI
14	Dong Y, Fan G, Li Y and Zhou Q: TUG1 represses apoptosis, autophagy, and inflammatory response by regulating miR-27a-3p/SLIT2 in LPS-treated vascular endothelial cells. J Surg Res. 256:345–354. 2020. View Article : Google Scholar : PubMed/NCBI
15	Li M, Wang J, Liu D and Huang H: High-throughput sequencing reveals differentially expressed lncRNAs and circRNAs, and their associated functional network, in human hypertrophic scars. Mol Med Rep. 18:5669–5682. 2018.PubMed/NCBI
16	Chen J, Zhou R, Liang Y, Fu X, Wang D and Wang C: Blockade of lncRNA-ASLNCS5088-enriched exosome generation in M2 macrophages by GW4869 dampens the effect of M2 macrophages on orchestrating fibroblast activation. FASEB J. 33:12200–12212. 2019. View Article : Google Scholar : PubMed/NCBI
17	Chen L and Li J, Li Q, Li X, Gao Y, Hua X, Zhou B and Li J: Overexpression of lncRNA AC067945.2 down-regulates collagen expression in skin fibroblasts and possibly correlates with the VEGF and Wnt signalling pathways. Cell Physiol Biochem. 45:761–771. 2018. View Article : Google Scholar : PubMed/NCBI
18	Tu L, Huang Q, Fu S and Liu D: Aberrantly expressed long noncoding RNAs in hypertrophic scar fibroblasts in vitro: A microarray study. Int J Mol Med. 41:1917–1930. 2018.PubMed/NCBI
19	Yamoto M, Alganabi M, Chusilp S, Lee D, Yazaki Y, Lee C, Li B and Pierro A: Lysosomal overloading and necrotizing enterocolitis. Pediatr Surg Int. 36:1157–1165. 2020. View Article : Google Scholar : PubMed/NCBI
20	Li Y, Zhang J, Zhou K, Xie L, Xiang G, Fang M, Han W, Wang X and Xiao J: Elevating sestrin2 attenuates endoplasmic reticulum stress and improves functional recovery through autophagy activation after spinal cord injury. Cell Biol Toxicol. Aug 1–2020.(Epub ahead of print). doi: 10.1007/s10565-020-09550-4. View Article : Google Scholar
21	Gutjahr A, Papagno L, Vernejoul F, Lioux T, Jospin F, Chanut B, Perouzel E, Rochereau N, Appay V, Verrier B and Paul S: New chimeric TLR7/NOD2 agonist is a potent adjuvant to induce mucosal immune responses. EBioMedicine. 58:1029222020. View Article : Google Scholar : PubMed/NCBI
22	Fu H, Chung KR, Liu X and Li H: Aaprb1, a subtilsin-like protease, required for autophagy and virulence of the tangerine pathotype of Alternaria alternata. Microbiol Res. 240:1265372020. View Article : Google Scholar : PubMed/NCBI
23	Shi J, Xiao H, Li J, Zhang J, Li Y, Zhang J, Wang X, Bai X, Tao K, Hu D and Guan H: Wild-type p53-modulated autophagy and autophagic fibroblast apoptosis inhibit hypertrophic scar formation. Lab Invest. 98:1423–1437. 2018. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Xu Q, Long X, Liu W, Zhao Y, Hayashi T, Hattori S, Fujisaki H, Ogura T, Tashiro SI, et al: Silibinin-induced autophagy mediated by PPARα-sirt1-AMPK pathway participated in the regulation of type I collagen-enhanced migration in murine 3T3-L1 preadipocytes. Mol Cell Biochem. 450:1–23. 2019. View Article : Google Scholar : PubMed/NCBI
25	Kyun S, Yoo C, Park HY, Kim J and Lim K: The effects of exogenous lactate administration on the IGF1/Akt/mTOR pathway in rat skeletal muscle. Int J Environ Res Public Health. 17:78052020. View Article : Google Scholar
26	Wang XC, Wang T, Zhang Y, Wang LL, Zhao RY and Tan W: Tacrolimus inhibits proliferation and induces apoptosis by decreasing survivin in scar fibroblasts after glaucoma surgery. Eur Rev Med Pharmacol Sci. 22:2934–2940. 2018.PubMed/NCBI
27	Jammal AA, Berchuck SI, Thompson AC, Costa VP and Medeiros FA: The effect of age on increasing susceptibility to retinal nerve fiber layer loss in glaucoma. Invest Ophthalmol Vis Sci. 61:82020. View Article : Google Scholar : PubMed/NCBI
28	Mu Y: Correlation analysis between TCM syndromes of primary angle-closure glaucoma and type A personality. Chin J Tradit Chin Ophthalmol. 10:23–30. 2020.
29	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
30	Ye Q, Wu J, Wu P, Wang J, Niu W, Yang S, Chen M, Rehman S and Zhu N: Enhancing peroxymonosulfate activation of Fe-Al layered double hydroxide by dissolved organic matter: Performance and mechanism. Water Res. 185:1162462020. View Article : Google Scholar : PubMed/NCBI
31	Wei Z, Wang Y, Jiang L, Ji N, Wang Y, Chen F, Li T, Li J, Xu H, Zeng X and Chen Q: miR-223 regulates oral squamous cell carcinoma metastasis through the Wnt/β-catenin signaling pathway. Oral Oncol. 109:1049412020. View Article : Google Scholar : PubMed/NCBI
32	Liu YP, Wan J, Long F, Tian J and Zhang C: circPVT1 facilitates invasion and metastasis by regulating miR-205-5p/c-FLIP axis in osteosarcoma. Cancer Manag Res. 12:1229–1240. 2020. View Article : Google Scholar : PubMed/NCBI
33	Zhang Y, Chang B, Zhang J and Wu X: lncRNA SOX2OT alleviates the high glucose-induced podocytes injury through autophagy induction by the miR-9/SIRT1 axis. Exp Mol Pathol. 110:1042832019. View Article : Google Scholar : PubMed/NCBI
34	Perez Vidakovics MLA, Ure AE, Arrias PN, Romanowski V and Gómez RM: Junin virus induces autophagy in human A549 cells. PLoS One. 14:e02187302019. View Article : Google Scholar : PubMed/NCBI
35	Neubert P, Weichselbaum A, Reitinger C, Schatz V, Schröder A, Ferdinand JR, Simon M, Bär AL, Brochhausen C, Gerlach RG, et al: HIF1A and NFAT5 coordinate Na⁺-boosted antibacterial defense via enhanced autophagy and autolysosomal targeting. Autophagy. 15:1899–1916. 2019. View Article : Google Scholar : PubMed/NCBI
36	Nong Q, Li S, Wu Y and Liu D: lncRNA COL1A2-AS1 inhibits the scar fibroblasts proliferation via regulating miR-21/Smad7 pathway. Biochem Biophys Res Commun. 495:319–324. 2018. View Article : Google Scholar : PubMed/NCBI
37	Wang X, Chu J, Wen CJ, Fu SB, Qian YL, Wo Y, Wang C and Wang DR: Functional characterization of TRAP1-like protein involved in modulating fibrotic processes mediated by TGF-β/Smad signaling in hypertrophic scar fibroblasts. Exp Cell Res. 332:202–211. 2015. View Article : Google Scholar : PubMed/NCBI
38	Pang K, Li B, Tang Z, Yang W, Hao L, Shi Z, Zhang J, Cai L, Li R, Liu Y, et al: Resveratrol inhibits hypertrophic scars formation by activating autophagy via the miR-4654/Rheb axis. Mol Med Rep. 22:3440–3452. 2020.PubMed/NCBI
39	Shi W, Wu Y and Bian D: p75NTR silencing inhibits proliferation, migration and extracellular matrix deposition of hypertrophic scar fibroblasts by activating autophagy through inhibiting PI3K/Akt/mTOR pathway. Can J Physiol Pharmacol. Jul 29–2020.(Epub ahead of print). doi: 10.1139/cjpp-2020-0219. View Article : Google Scholar
40	De Risi M, Torromino G, Tufano M, Moriceau S, Pignataro A, Rivagorda M, Carrano N, Middei S, Settembre C, Ammassari-Teule M, et al: Mechanisms by which autophagy regulates memory capacity in ageing. Aging Cell. 19:e131892020. View Article : Google Scholar : PubMed/NCBI
41	Lanzillotta C, Zuliani I, Vasavda C, Snyder SH, Paul BD, Perluigi M, Di Domenico F and Barone E: BVR-A deficiency leads to autophagy impairment through the dysregulation of AMPK/mTOR axis in the brain-implications for neurodegeneration. Antioxidants (Basel). 9:6712020. View Article : Google Scholar
42	Long J and Pi X: Polyphyllin I promoted melanoma cells autophagy and apoptosis via PI3K/Akt/mTOR signaling pathway. Biomed Res Int. 2020:51494172020. View Article : Google Scholar : PubMed/NCBI
43	Luo M, Liu Y, Liu N, Shao W, Ming L, Liu J and Xie Y: Proscillaridin A inhibits hepatocellular carcinoma progression through inducing mitochondrial damage and autophagy. Acta Biochim Biophys Sin (Shanghai). Nov 17–2020.(Epub ahead of print). doi: 10.1093/abbs/gmaa139. View Article : Google Scholar
44	Yang J, Wang B, Xu Q, Yang Y, Hou L, Yin K, Guo Q, Hua Y, Zhang L, Li Y, et al: TMEM166 inhibits cell proliferation, migration and invasion in hepatocellular carcinoma via upregulating TP53. Mol Cell Biochem. Nov 16–2020.(Epub ahead of print). doi: 10.1007/s11010-020-03979-1. View Article : Google Scholar
45	Yang Z, Tang Y, Lu H, Shi B, Ye Y, Xu G and Zhao Q: Long non-coding RNA reprogramming (lncRNA-ROR) regulates cell apoptosis and autophagy in chondrocytes. J Cell Biochem. 119:8432–8440. 2018. View Article : Google Scholar : PubMed/NCBI
46	Urut F, Dede S, Yuksek V, Cetin S, Usta A and Taspinar M: In vitro evaluation of the apoptotic, autophagic, and necrotic molecular pathways of fluoride. Biol Trace Elem Res. Nov 16–2020.(Epub ahead of print). doi: 10.1007/s12011-020-02491-3. View Article : Google Scholar : PubMed/NCBI