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LeftyA sensitive cytosolic pH regulation and glycolytic flux in Ishikawa human endometrial cancer cells.

Abstract：

OBJECTIVE:LeftyA, a powerful regulator of stemness, embryonic differentiation, and reprogramming of cancer cells, counteracts cell proliferation and tumor growth. Key properties of tumor cells include enhanced glycolytic flux, which is highly sensitive to cytosolic pH and thus requires export of H(+) and lactate. H(+) extrusion is in part accomplished by Na(+)/H(+) exchangers, such as NHE1. An effect of LeftyA on transport processes has, however, never been reported. The present study thus explored whether LeftyA modifies regulation of cytosolic pH (pHi) in Ishikawa cells, a well differentiated endometrial carcinoma cell model. METHODS:NHE1 transcript levels were determined by qRT-PCR, NHE1 protein abundance quantified by Western blotting, pHi estimated utilizing (2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein [BCECF] fluorescence, Na(+)/H(+) exchanger activity from Na(+) dependent realkalinization after an ammonium pulse, and lactate concentration in the supernatant utilizing an enzymatic assay and subsequent colorimetry. RESULTS:A 2 h treatment with LeftyA (8 ng/ml) significantly decreased NHE1 transcript levels (by 99.6%), NHE1 protein abundance (by 71%), Na(+)/H(+) exchanger activity (by 55%), pHi (from 7.22 ± 0.02 to 7.05 ± 0.02), and lactate release (by 41%). CONCLUSIONS:LeftyA markedly down-regulates NHE1 expression, Na(+)/H(+) exchanger activity, pHi, and lactate release in Ishikawa cells. Those effects presumably contribute to cellular reprogramming and growth inhibition.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Salker MS,Zhou Y,Singh Y,Brosens J,Lang F

doi

10.1016/j.bbrc.2015.03.120

subject

Has Abstract

pub_date

2015-05-08 00:00:00

pages

845-9

issue

3

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(15)00586-0

journal_volume

460

pub_type

杂志文章

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