Mass Spectrometry Reveals α-2-HS-Glycoprotein as a Key Early Extracellular Matrix Protein for Conjunctival Cells

Abstract

Purpose To determine the composition of extracellular matrix (ECM) proteins secreted by a conjunctival epithelial cell line and to identify components that aid conjunctival epithelial cell culture. Methods Human conjunctival epithelial cell line (HCjE-Gi) cells were cultured in serum-free media and their ECM isolated using ammonium hydroxide. Growth characteristics were evaluated for fresh HCjE-Gi cells plated onto ECMs obtained from 3- to 28-day cell cultures. Mass spectrometry was used to characterize the ECM composition over 42 culture days. Cell adhesion and growth on pre-adsorbed fibronectin and α-2-HS-glycoprotein (α-2-HS-GP) were investigated. Results Day 3 ECM provided the best substrate for cell growth compared to ECM obtained from 5- to 28-day cell cultures. Mass spectrometry identified a predominantly laminin 332 matrix throughout the time course, with progressive changes to matrix composition over time: proportional decreases in matrix-bound growth factors and increases in proteases. Fibronectin and α-2-HS-GP were 5- and 200-fold enriched as a proportion of the early ECM relative to the late ECM, respectively. Experiments on these proteins in isolation demonstrated that fibronectin supported rapid cell adhesion, whereas fibronectin and α-2-HS-GP both supported enhanced cell growth compared to tissue culture polystyrene. Conclusions These data reveal α-2-HS-GP as a candidate protein to enhance the growth of conjunctival epithelial cells and raise the possibility of exploiting these findings for targeted improvement to synthetic tissue engineered conjunctival substrates.

Alpha‐2‐Heremans‐Schmid‐glycoprotein (AHSG) a potential biomarker associated with prognosis of chromophobe renal cell carcinoma: The PROPOLIS study

“…According to further analysis of the gene expression between the dead chRCC patients ( n = 9) and the surviving chRCC patients ( n = 56) (Figure 9A), only the AHSG gene survived significantly less in chRCC contrasted to normal samples ( p < 0.05) (Figure 8), suggesting that AHSG may be a potential biomarker associated with prognosis of chRCC. The top 20 proteins interacting with AHSG by GeneMANIA were partially associated with tumor development or tumor suppression, which also suggests that AHSG is a potential key gene for chRCC 26,27,29,30,33–35,37,38 …”

“…The result showed that the top 20 proteins interacting with AHSG (Figure 9B ) mainly functioned in tumor suppressor, signal transduce, cancer development. 26 , 27 , 29 , 30 , 33 , 34 , 35 , 36 , 37 , 38 , 39…”

“…The top 20 proteins interacting with AHSG by GeneMANIA were partially associated with tumor development or tumor suppression, which also suggests that AHSG is a potential key gene for chRCC. 26 , 27 , 29 , 30 , 33 , 34 , 35 , 37 , 38 …”

“…The result showed that the top 20 proteins interacting with AHSG (Figure 9B) mainly functioned in tumor suppressor, signal transduce, cancer development. 26,27,29,30,[33][34][35][36][37][38][39] 4 | DISCUSSION ccRCC is a malignant tumor originating from the collecting epithelial cells of the renal tubules, which has no obvious clinical symptoms in the early stage and is clinically uncommon. However, some patients still have a poor prognosis and patients with the advanced disease require adjuvant therapy.…”

The conjunctival extracellular matrix, related disorders and development of substrates for conjunctival restoration

“…The BM is a dynamic structure the composition of which changes not only during development, but also with age and following repair of injury. Although not measured in vivo, in vitro studies using a human immortalized conjunctival cell line (HCjE) showed changes to the composition of the ECM over a twenty-eight day culture period (54). Using mass spectrometry, the whole proteome of the ECM was studied and proteins such as -2-HS-glycoprotein were identified in the conjunctival ECM.…”