Alterations in atp-dependence ofswelling- activated cl- currentassociated with neuroendocrinedifferentiation of lncap prostatecancer epithelial cells
R.M. Lazarenko, A.P. KondratskyN.Ch. Pogorela, Y.M. Shuba
A.A.Bogomoletz Institute of Physiology, National Academy ofScience of Ukraine, Kyiv
Increasing population of malignant, apoptosis resistant
neuroendocrine (NE) cells due to differentiation of prostate
epithelial/basal cells is a hallmark of advanced, androgen-inde-
pendent prostate cancer, for which there is no successful
therapy. Acquisition of apoptosis resistance involves alter-
ations in the mechanisms of cell volume homeostasis, of which
volume-regulate anion channels (VRAC) that carry swelling-
activated Cl- current (ICl,swell) represent one of the key determi-
nants. Given that VRAC function is generally known to be
ATP-dependent, here we investigated how such dependence
may evolve during NE differentiation of LNCaP prostate cancer
epithelial cells. In the whole-cell patch-clamp recording mode
ICl,swell could be activated in response to hypotonicity-induced
cell swelling in control and NE-differentiated (by incubation in
membrane-permeable cAMP analogs) LNCaP cells even fol-
lowing total depletion of intracellular ATP using a cocktail of
metabolic inhibitors. However, this basal ICl,swell had about
30% higher density and was less inactivating in NE-differenti-
ated cells. Inclusion of 5 mM Mg-ATP in the patch pipette
caused ICl,swell augmentation in both cell types. The augmenta-
tion in the control cells was more prominent and occurred
mostly at the expense of a non-inactivating current component.
We conclude that ICl,swell in LNCaP cells consists of a non-
inactivating, ATP-dependent and inactivating, ATP-indepen-
dent components. NE differentiation promotes the increase of
non-inactivating component and partial loss of its ATP sensi-
tivity making the whole ICl,swell less ATP-sensitive as well. By
largely avoiding the ATP metabolic control ICl,swell may contrib-
ute to better control of cell volume under metabolic stress and
thus enhance the survival rates of apoptosis-resistant NE cells.
- Aprikian A.G., Cordon-Cardo C., Fair W.R., ReuterV.E. Characterization of neuroendocrine differentiationin human benign prostate and prostatic adenocarcinoma// Cancer. – 1993. – 71(12). – P. 3952–3965.
- Bond T., Basavappa S., Christensen M., Strange K.ATP Dependence of the ICl,swell Channel Varies withthe Rate of Cell Swelling // J. Gen. Physiol. – 1999. –113. – P. 441–456.
- Bonkhoff H. Neuroendocrine differentiation in humanprostate cancer. Morphogenesis, proliferation and an-drogen receptor status // Ann. Oncol. – 2001. – 12. –P. 141–144.
- Cox M.E., Deeble P.D., Bissonette E.A., Parsons S.J.Activated 3',5'-cyclic AMP-dependent protein kinaseis sufficient to induce neuroendocrine-like differentia-tion of the LNCaP prostate tumor cell line // J. Biol.Chem. – 2000. – 275(18). – P. 13812–13818.
- Cox M.E., Deeble P.D., Lakhani S., Parsons S.J. Acquisi-tion of neuroendocrine characteristics by prostate tumorcells is reversible: implications for prostate cancer pro-gression // Cancer Res. – 1999. – 59. – P. 3821–3830.
- Culic O., Gruwel M.L., Schrader J. Energy turnover ofvascular endothelial cells // Amer. J. Physiol. – 1997. –273. – P. 205–213.
- Eggermont J., Trouet D., Carton I., Nilius B. Cellularfunction and control of volume-regulated anionchannels // Cell Biochem. and Biophys. – 2001. – 35. –P. 263–274.
- Geschwind J.–F.H., Ko Y.H., Torbenson M.S. et al.Novel therapy for liver cancer: direct intraarterial in-jection of a potent inhibitor of ATP production // CancerRes. – 2002. – 62. – P. 3909–3913.
- 9. Golenhofen N., Doctro R.B., Baccalao R., Mandell L.J.and villin compartmentation during ATP deple-tion and recovery in renal cultured cells // Kidney Int. –1996. – 48. – P. 1837–1845.
- 10. Ito T., Yamamoto S., Ohno Y. et al. Up-regulation ofneuroendocrine differentiation in prostate cancer afterandrogen deprivation therapy, degree and androgenindependence // Oncol. Rep. – 2001. – 8. – P. 1221–1224.
- Kim J.H., Shin S.Y., Yun S.S. et al . Voltage-dependention channel currents in putative neuroendocrine cellsdissociated from the ventral prostate of rat // Pflug.Arch. – 2003. – 446 (1). – P. 88–89.
- Krijnen J.L., Janssen P.J., Ruizeveld de Winter J.A. etal. Do neuroendocrine cells in human prostate cancerexpress androgen receptor? // Histochemistry. – 1993. –100. – P. 393–398.
- Lehninger A.L. Principles of Biochemistry // WorthPublishers, Inc. – 1985. – 2. – P. 385–551.
- Lemonnier L., Lazarenko R., Shuba Y. et al. Alterationsregulatory volume decrease and swelling–activatedCl- current associated with neuroendocrine differen-tiation of prostate cancer epithelial cells // Endocr. Relat.Cancer. – 2005. – 12(2). – P. 335-249.
- Lemonnier L., Prevarskaya N., Shuba Y. et al. Ca2+modulation of volume–regulated anion channels: evi-dence for colocalization with store–operated channels// FASEB J. – 2002. – 16. – P. 222–224.
- Lemonnier L., Shuba Y., Crepin A. et al . Bcl-2-depen-dent modulation of swelling-activated C- current andClC-3 expression in human prostate cancer epithelialcells // Cancer Res. :4841–8. – 2004. – 64(14). –P. 4841–4848.
- Li Y.C., Fung K.P., Kwok T.T. et al. Mitochondria-targeting drug oligomycin blocked P-glycoprotein ac-tivity and triggered apoptosis in doxorubicin–resistantHepG2 cells // Chemotherapy. – 2004. – 50. – P. 55–62.
- Martin D.S., Bertino J.R., Koutcher J.A. ATP deple-tion + pyrimidine depletion can markedly enchancecancer therapy: fresh insight for a new approach //Cancer Res. – 2000. – 60. – P. 6776–6783.
- 19. Mills J.W., Lubin M. Effect of adenosine 3',5'-cyclicmonophosphate on volume and cytoskeleton ofMDCK cells // Amer. J. Physiol. – 1986. – 250 (2 Pt1). – P. C319–324.
- 20. Nieminen A.L. Apoptosis and necrosis in health anddisease: role of mitochondria // Int. Rev. Cytol. – 2003. –224. – P. 29–55.
- Okada Y. Volume expansion-sensing outward-rectifierCl- channel: fresh start to the molecular identity andvolume sensor // Amer. J. Physiol. – 1997. – 273 (3 Pt 1). –P.755–789.
- Shen M.R., Yang T.P., Tang M.J. A novel function ofBCL-2 overexpression in regulatory volume decrease.Enhancing swelling–activated Ca2+ entry and Cl?channel activity // J. Biol. Chem. – 2002. – 277. –P. 15592–15599.
- Shuba Y.M., Prevarskaya N., Lemonnier L. et al. Vol-ume-regulated chloride conductance in the LNCaP hu-man prostate cancer cell line // Amer. J. Physiol. –2000. – 279. – P. C1144–C1154
- Tai K.K., McCrossan Z.A., Abbott G.W. Activationof mitochondrial ATP-sensitive potassium channelsincreases cell viability against rotenone-induced celldeath // J. Neurochem. – 2003. – 84. – P. 1193–2000.
- Vashchenko N., Abrahamson P.-A. Neuroendocrinedifferentiation in prostate cancer: Implication for newtreatment modalities // Europ. Urol. – 2005. – 47. –P. 147–155.