Biomedical subjects

Shiwei Cai

Publications and source records attributed to Shiwei Cai.

3 recordsLinked to original sources

PubMed2005

Evidence that TRPC1 contributes to calcium-induced differentiation of human keratinocytes.

External calcium ion concentration is a major regulator of epidermal keratinocyte differentiation in vitro and probably also in vivo. Regulation of calcium-induced differentiation changes is proposed to occur via an external calcium-sensing, signaling pathway that utilizes increases in intracellular calcium ion concentration to activate differentiation-related gene expression. Calcium ion release from intracellular stores and calcium ion influx via store-operated calcium-permeable channels are key elements in this proposed signaling pathway; however, the channels involved have not yet been identified. The present report shows that human gingival keratinocytes (HGKs) also undergo calcium-induced differentiation in vitro as indicated by involucrin expression and morphological changes. Moreover, TRPC1, which functions as a store-operated calcium channel in a number of cell types, including epidermal keratinocytes, is expressed in both proliferating and differentiating HGKs. Transfection of HGKs with TRPC1 siRNA disrupted expression of TRPC1 mRNA and protein compared with transfection with scrambled TRPC1 siRNA. Cells with disrupted TRPC1 expression showed decreased calcium-induced differentiation as measured by involucrin expression or morphological changes, as well as decreased thapsigargin-induced calcium ion influx, and a decreased rate of store calcium release. These results indicate that TRPC1 is involved in calcium-induced differentiation of HGKs likely by supporting a store-operated calcium ion influx.

Calcium↗

PubMed2005

TRPC channel expression during calcium-induced differentiation of human gingival keratinocytes.

BACKGROUND: Extracellular calcium is an important regulator of keratinocyte differentiation. An increase in intracellular calcium ion concentration is required for activation of calcium-induced keratinocyte differentiation. The signaling elements in this differentiation response include the calcium sensing receptor, phospholipase C, release of calcium ions from intracellular stores, and store-operated calcium channels. Nothing is currently known about the calcium-entry channels activated by the increase in external calcium. However, canonical transient receptor potential (TRPC) channels have been identified as store-operated calcium channels in several tissues. OBJECTIVE: To examine the expression of TRPC channels in human gingival keratinocytes (HGKs) in primary culture under both low calcium (basal) and high calcium (differentiating) conditions, and in gingival tissue. METHODS: TRPC channel expression was evaluated via RT-PCR, Western blots, and immunohistology. RESULTS: TRPC1, TRPC5, TRPC6 and TRPC7 mRNAs were detected in undifferentiated keratinocytes. Their levels initially increased, then decreased during calcium-induced differentiation. TRPC1 and TRPC6 protein expression reflected these changes. CONCLUSION: TRPC channels are present in both proliferating and differentiating keratinocytes in primary culture and in gingival tissue. The above expression patterns suggest that these channels may be involved in calcium-induced differentiation of keratinocytes.

Calcium↗

PubMed2004

Calcium receptor message, expression and function decrease in differentiating keratinocytes.

Calcium-sensing receptor (CaSR) expression and function were studied in proliferating and differentiating cultured human gingival keratinocytes (HGKs). CaSR mRNA and protein were present in proliferating HGKs cultured in 0.03 mM [Ca(2+)] and decreased in cells induced to differentiate by culturing in 1.2 mM [Ca(2+)] for 2 days. CaSR protein was also detected in gingival tissue. Exposure to 10 mM extracellular [Ca(2+)] activated two sequential whole-cell currents. The first was a small, transient calcium release activated calcium current I(CRAC)-like current with an inwardly rectifying I-V curve. The second current was larger with a linear I-V curve. Both currents were significantly decreased in differentiating cells. Neither neomycin nor gadolinium induced changes in whole cell currents nor in intracellular [Ca(2+)], but neomycin inhibited the late large current. Extracellular Ca(2+) increased intracellular [Ca(2+)] of proliferating HGKs in a dose-dependent fashion. Comparison of the time-courses of the whole-cell currents and the intracellular [Ca(2+)] responses indicated both induced currents supported a Ca(2+) influx. Extracellular [Mg(2+)] changes did not affect intracellular [Ca(2+)]. La(3+) and 2-APB inhibited the whole cell current and intracellular [Ca(2+)] changes. The results indicate that the CaSR signaling response likely plays a major role in initiating Ca(2+) induced differentiation responses in HGKs.

Blotting, Western↗