Xiuxiang LIU, ZHICHUN FENG, Chunmei TIAN, Xiangyong KONG, YUMEI WU, Changjun LV

Effect of keratinocyte growth factor on growth and transdifferentiation of primary alveolar epithelial type II cells

Background/aim: To investigate the effects of keratinocyte growth factor (KGF) on the growth and transdifferentiation of primary alveolar epithelial type II cells (AECIIs). Materials and methods: The number of primary AECIIs, their viability, and cell cycle and apoptosis were studied under KGF treatment using a hemocytometer, trypan blue exclusion, and flow cytometry, respectively. Positive expressions of surfactant protein C (SP-C), aquaporin 5 (AQP5), and thyroid transcription factor 1(TTF-1) were examined with indirect immunofluorescence; mRNA levels of SP-C, AQP5, and TTF-1 were determined by polymerase chain reaction. Results: In response to KGF treatment, cell numbers were significantly increased, more cells were blocked in the S phase, and fewer cells were apoptotic or necrotic on days 2 and 4, but there was little effect on cell viability. In addition, KGF treatment resulted in higher levels of SP-C on days 2 and 8 while lowering them on day 4, higher levels of AQP5 on day 4 while lowering them on day 8, and higher levels of TTF-1 on days 2, 6, and 8. Conclusion: KGF treatment promoted proliferation of AECIIs, inhibited cell apoptosis, promoted transdifferentiation of AECIIs, and induced alveolar epithelial type I cells to revert to AECIIs.

Anahtar Kelimeler:

Keratinocyte growth factor, alveolar epithelial type II cells, transdifferentiation, surfactant protein C, aquaporin 5, thyroid transcription factor-1

Effect of keratinocyte growth factor on growth and transdifferentiation of primary alveolar epithelial type II cells

Keywords:

Keratinocyte growth factor, alveolar epithelial type II cells, transdifferentiation, surfactant protein C, aquaporin 5, thyroid transcription factor-1,

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Pottier N, Maurin T, Chevalier B, Puisségur MP, Lebrigand K, Robbe-Sermesant K, Bertero T, Lino Cardenas CL, Courcot E, Rios G et al. Identification of keratinocyte growth factor as a target of microRNA-155 in lung fibroblasts: implication in epithelial-mesenchymal interactions. PLoS One 2009; 4: e6718.
LaFemina MJ, Rokkam D, Chandrasena A, Pan J, Bajaj A, Johnson M, Frank JA. Keratinocyte growth factor enhances barrier function without altering claudin expression in primary alveolar epithelial cells. Am J Physiol Lung Cell Mol Physiol 2010; 299: L724–L734.
She J, Goolaerts A, Shen J, Bi J, Tong L, Gao L, Song Y, Bai C. KGF-2 targets alveolar epithelia and capillary endothelia to reduce high altitude pulmonary oedema in rats. J Cell Mol Med 2012; 16: 3074–3084.
Crosby LM, Waters CM. Epithelial repair mechanisms in the lung. Am J Physiol Lung Cell Mol Physiol 2010; 298: L715–731.
Saada J, Oudrhiri N, Bonnard A, de Lagausie P, Aissaoui A, Hauchecorne M, Oury JF, Aigrain Y, Peuchmaur M, Lehn JM et al. Combining keratinocyte growth factor transfection into the airways and tracheal occlusion in a fetal sheep model of congenital diaphragmatic hernia. J Gene Med 2010; 12: 413– 422.
Qiao R, Yan W, Clavijo C, Mehrian-Shai R, Zhong Q, Kim KJ, Ann D, Crandall ED, Borok Z. Effects of KGF on alveolar epithelial cell transdifferentiation are mediated by JNK signaling. Am J Respir Cell Mol Biol 2008; 38: 239–246.
Cheng CC, Wang DY, Kao MH, Chen JK. The growth- promoting effect of KGF on limbal epithelial cells is mediated by upregulation of ΔNp63α through the p38 pathway. J Cell Sci 2009; 122: 4473–4480.
Liu XX, Yu XR, Jia XH, Wang KX, Yu ZY, Lv CJ. Effect of hyperoxia on the viability and proliferation of the primary type II alveolar epithelial cells. Cell Biochem Biophys 2013; 67: 1539–1546. 9. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method. Methods 2001; 25: 402–408.
Wang D, Haviland DL, Burns AR, Zsigmond E, Wetsel RA. A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells. P Natl Acad Sci USA 2007; 104: 4449–4454.
Das A, Acharya S, Gottipati KR, McKnight JB, Chandru H, Alcorn JL, Boggaram V. Thyroid transcription factor-1 (TTF- 1) gene: identification of ZBP-89, Sp1, and TTF-1 sites in the promoter and regulation by TNF-α in lung epithelial cells. Am J Physiol Lung Cell Mol Physiol 2011; 301: L427–L440.
Ishikawa A, Kudo M, Nakazawa N, Onda M, Ishiwata T, Takeshita T, Naito Z. Expression of keratinocyte growth factor and its receptor in human endometrial cancer in cooperation with steroid hormones. Int J Oncol 2008; 32: 565–574.
Gomperts BN, Belperio JA, Fishbein MC, Keane MP, Burdick MD, Strieter RM. Keratinocyte growth factor improves repair in the injured tracheal epithelium. Am J Respir Cell Mol Biol 2007; 37: 48–56.
Liu XX, Lv CJ, Jia XH, Feng ZC. A systemic study on the protective effect of keratinocyte growth factor on type II alveolar epithelial cells against hyperoxia-induced injury in vitro. Health Med 2009; 3: 225–233.
Sadovski J, Kuchenbuch T, Ruppert C, Fehrenbach A, Hirschburger M, Padberg W, Günther A, Hohlfeld JM, Fehrenbach H, Grau V. Keratinocyte growth factor prevents intra-alveolar oedema in experimental lung isografts. Eur Respir J 2008; 31: 21–28.
Baybutt RC, Smith BW, Donskaya EV, Hu L, Li T, Wang W. The proliferative effects of retinoic acid on primary cultures of adult rat type II pneumocytes depend upon cell density. In Vitro Cell Dev Biol Anim 2010; 46: 20–27.
Yen TT, Thao DT, Thuoc TL. An overview on keratinocyte growth factor: from the molecular properties to clinical applications. Protein Pept Lett 2014; 21: 306–317.
Liu XX, Zhang HH, Wan B, Luo Y, Feng ZC. Growing conditions of type II alveolar epithelial cells and their transdifferent identification in vitro. Journal of Applied Clinical Pediatrics 2011; 26: 1287–1291 (in Chinese with abstract in English).
Lin J, Liu C, Ge L, Gao Q, He X, Liu Y, Li S, Zhou M, Chen Q, Zhou H. Carcinoma-associated fibroblasts promotes the proliferation of a lingual carcinoma cell line by secreting keratinocyte growth factor. Tumour Biol 2011; 32: 597–602.
Mei C, Mao Z, Shen X, Wang W, Dai B, Tang B, Wu Y, Cao Y, Zhang S, Zhao H et al. Role of keratinocyte growth factor in the pathogenesis of autosomal dominant polycystic kidney disease. Nephrol Dial Transplant 2005; 20: 2368–2375.
Wildhaber BE, Yang H, Teitelbaum DH. Keratinocyte growth factor decreases total parenteral nutrition-induced apoptosis in mouse intestinal epithelium Via Bcl-2. J Pediatr Surg 2003; 38: 92–96.
Wildhaber BE, Yang H, Teitelbaum DH. Total parenteral nutrition-induced apoptosis in mouse intestinal epithelium: modulation by keratinocyte growth factor. J Surg Res 2003; 112: 144–151.
Runkle EA, Rice SJ, Qi J, Masser D, Antonetti DA, Winslow MM, Mu D. Occludin is a direct target of thyroid transcription factor-1 (TTF-1/NKX2-1). J Biol Chem 2012; 287: 28790– 28801.
Tichelaar JW, Lu W, Whitsett JA. Conditional expression of fibroblast growth factor-7 in the developing and mature lung. J Biol Chem 2000; 275: 11858–11864.