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Modulation of volume-sensitive Cl – channels and cell volume by actin filaments and microtubules in human cervical cancer HT-3 cells

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Hypotonicity activates volume-sensitive Cl currents, which are implicated in the regulatory volume decrease (RVD) responses and transport of taurine in human cervical cancer HT-3 cells. In this study, the role of cytoskeleton in the regulation of volume-sensitive Cl channels and RVD responses in HT-3 cells was studied. Cells were incubated with various compounds, which depolymerized or polymerized cytoskeletal elements, i.e. actin filaments and microtubules. The hypotonicity-induced changes in Cl conductance and in cell volume were measured by whole-cell voltage clamping and cell size monitoring, respectively. Our results show that in HT-3 cells hypotonicity activated an outward rectified Cl current that was abrogated by Cl channel blockers. Cytochalasin B, an actin-depolymerizing compound, induced a substantial increase in Cl conductance under isotonic condition and potentiated the expression of Cl currents in hypotonic stress. Phorbol 12-myristate 13-acetate (PMA) significantly inhibited the cytochalasin B-induced activation of Cl conductance under isotonic condition. On the other hand, treatment with cytochalasin B significantly prolonged the RVD responses. Phalloidin, a stabilizer of actin polymerization, did not change the basal currents under isotonic condition, but completely abolished the increase in whole-cell Cl conductance elicited by hypotonicity and retarded the cell volume recovery. Colchicine, a microtubule-assembly inhibitor, had no effect on either basal Cl conductance or volume-sensitive Cl current and was unable to inhibit the RVD responses. Taxol, a microtubule-stabilizing compound, did not alter the basal Cl conductance, but inhibited the activation of volume-sensitive Cl channels as well as the process of RVD in a dose-dependent manner. These data support the notion that functional integrity of actin filaments and microtubules plays critical roles in maintaining the RVD responses and activation of Cl channels in human cervical cancer HT-3 cells.

Keywords: actin; cell-volume regulation; microtubule; regulatory volume decrease; volume-sensitive Cl– channels

Document Type: Research Article


Affiliations: 1: Department of Obstetrics and Gynecology, National Cheng Kung University Medical College, Tainan, Taiwan 2: Department of Pharmacology, National Cheng Kung University Medical College, Tainan, Taiwan

Publication date: 1999-11-01

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