Down-regulation of lncRNA-H19 expression promotes apoptosis and autophagy of human keloid fibroblasts

doi:10.16352/j.issn.1001-6325.2022.04.023

Abstract

Abstract: Objective To investigate the effects of down-regulation of long non-coding RNA-H19 (lncRNA-H19) expression on apoptosis and autophagy of human keloid fibroblast cell line and to explore potential mechanism. Methods Human scar fibroblasts cell line (KFs) and normal human dermal fibroblasts cell line (HDFs) were collected and KFs cells were infected with lncRNA-H19F low-expressing adenovirus (si-lncRNA-H19) and empty virus vector without si-lncRNA-H19 (si-eGFP). They were divided into HDFs group, KFs group, si-lncRNA-H19+KFs group and si-eGFP+KFs group. KFs cells were co-transfected with si-lncRNA, mTOR over-expressing adenovirus (Ad-mTOR) and empty vector (Ad-eGFP-m), then divided into KFs group, si-lncRNA-H19+Ad-mTOR group,si-eGFP+Ad-mTOR group, si-lncRNA-H19+Ad-eGFP group and si-eGFP+Ad-eGFP group. RT-qPCR was used to detect the expression of lncRNA-H19; flow cytometry was used to detect the cell apoptosis ; autophagy was examined by EM microscopy with fluorescence (acridine orange,AO) staining; immuno-histochemical method was used to detect the positive expression of autophagy pathway-mammalian target of rapamycin (mTOR); Western blot was used to detect the expression of apoptosis and autophagy-related proteins. Results Compared with the HDFs group, the expression of lncRNA-H19 and mTOR activity in the KFs group increased and the number of autophagic vesicles and the expression of apoptosis and autophagy-related proteins decreased (P＜0.05); after down-regulating lncRNA-H19 in KFs cells, the activity of mTOR in KFs cells decreased and apoptosis/autophagy increased (P＜0.05). Up-regulating mTOR while down-regulating lncRNA-H19 lead to more autophagy and apoptotic in KFs cells as compared to that resulted from down-regulation of lncRNA-H19 alone (P＜0.05). Conclusions Down-regulation of lncRNA-H19 expression may promote the autophagy and apoptosis of KFs cells through inhibiting activation of mTOR pathway.

Key words: long non-coding H-19, human keloid fibroblasts, apoptosis, autophagy

CLC Number:

R622

HUANG Yun, XU Xi-sheng, CHEN Kai, HE Xiu. Down-regulation of lncRNA-H19 expression promotes apoptosis and autophagy of human keloid fibroblasts[J]. Basic & Clinical Medicine, 2022, 42(4): 633-639.

References

[1] 郭亚东, 吴红娟, 孙要文. PODXL参与瘢痕疙瘩成纤维细胞的过度增殖[J]. 基础医学与临床, 2019, 39:1243-1247.
[2] Magni G, Banchelli M, Cherchi F, et al. Experimental study on blue light interaction with human keloid-derived fibroblasts[J]. Biomedicines,2020,8:573-578.
[3] Si L, Zhang M, Guan E, et al. Resveratrol inhibits proliferation and promotes apoptosis of keloid fibroblasts by targeting HIF-1α[J]. J Plast Surg Hand Surg,2020,54:290-296.
[4] Sun CM, Enkhjargal B, Reis C, et al. Osteopontin attenuates early brain injury through regulating autophagy-apoptosis interaction after subarachnoid hemorrhage in rats[J]. CNS Neurosci Ther,2019,25:1162-1172.
[5] Chen K, Yang D, Zhao F, et al. Autophagy and tumor database: ATdb, a novel database connecting autophagy and tumor[J]. Database (Oxford), 2020, 20:52-58.
[6] Wang Q, Wang P, Qin Z, et al. Altered glucose metabolism and cell function in keloid fibroblasts under hypoxia[J]. Redox Biol,2021,38:1018-1025.
[7] Simion V, Zhou H, Haemmig S, et al. A macrophage-specific lncRNA regulates apoptosis and atherosclerosis by tethering HuR in the nucleus[J]. Nat Commun,2020,11:6135-6146.
[8] Wang Z, Feng C, Song K, et al. lncRNA-H19/miR-29a axis affected the viability and apoptosis of keloid fibro-blasts through acting upon COL1A1 signaling[J]. J Cell Biochem, 2020,121:4364-4376.
[9] Kouotou EA, Nansseu JR, Omona Guissana E, et al. Epidemiology and clinical features of keloids in Black Africans: a nested case-control study from Yaoundé, Came-roon[J]. Int J Dermatol,2019,58:1135-1140.
[10] Tan T, Li J, Wen Y, et al. Association between lncRNA-H19 polymorphisms and hepatoblastoma risk in an ethic Chinese population[J]. J Cell Mol Med, 2020, 25:742-750.
[11] Xu L, Sun N, Li G, et al. LncRNA H19 promotes keloid formation through targeting the miR-769-5p/EIF3A pathway[J]. Mol Cell Biochem,2021,3:1020-1024.
[12] Bhardwaj N, Tripathi N, Goel B, et al. Anticancer activity of diosgenin and its semi-synthetic derivatives: role in autophagy mediated cell death and induction of apoptosis[J]. Mini Rev Med Chem, 2021,4:2174-2185.
[13] Cui C, Han S, Tang S, et al. The autophagy regulatory molecule CSRP3 interacts with LC3 and protects against muscular dystrophy[J]. Int J Mol Sci, 2020,21:749-752.
[14] Tang Z, Cao Y, Ding J, et al. Wubeizi ointment suppresses keloid formation through modulation of the mTOR pathway[J]. Biomed Res Int, 2020,30:3608-3612.
[15] Huang TJ, Ren JJ, Zhang QQ, et al. IGFBPrP1 accelerates autophagy and activation of hepatic stellate cells via mutual regulation between H19 and PI3K/AKT/mTOR pathway[J]. Biomed Pharmacother, 2019,116:1093-1099.