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TRPV6 determines the effect of vitamin D3 on prostate cancer cell growth.

Lehen'kyi V, Raphaël M, Oulidi A, Flourakis M, Khalimonchyk S, Kondratskyi A, Gordienko DV, Mauroy B, Bonnal JL, Skryma R, Prevarskaya N - PLoS ONE (2011)

Bottom Line: We show that these pro-proliferative effects of 1,25-dihydroxyvitamin D3 are directly mediated via the overexpression of TRPV6 channel which increases calcium uptake into LNCaP cells.The apoptosis resistance of androgen-dependent LNCaP cells conferred by TRPV6 channel is drastically inversed when 1,25-dihydroxyvitamin D3 effects were combined with the successful TRPV6 knockdown.In addition, the use of androgen-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines allowed to suggest that the ability of 1,25-dihydroxyvitamin D3 to induce the expression of TRPV6 channel is a crucial determinant of the success or failure of 1,25-dihydroxyvitamin D3-based therapies.

View Article: PubMed Central - PubMed

Affiliation: Inserm, U-1003, Equipe labellisée par la Ligue Nationale contre le cancer, Villeneuve d'Ascq, France.

ABSTRACT
Despite remarkable advances in the therapy and prevention of prostate cancer it is still the second cause of death from cancer in industrialized countries. Many therapies initially shown to be beneficial for the patients were abandoned due to the high drug resistance and the evolution rate of the tumors. One of the prospective therapeutical agents even used in the first stage clinical trials, 1,25-dihydroxyvitamin D3, was shown to be either unpredictable or inefficient in many cases. We have already shown that TRPV6 calcium channel, which is the direct target of 1,25-dihydroxyvitamin D3 receptor, positively controls prostate cancer proliferation and apoptosis resistance (Lehen'kyi et al., Oncogene, 2007). However, how the known 1,25-dihydroxyvitamin D3 antiproliferative effects may be compatible with the upregulation of pro-oncogenic TRPV6 channel remains a mystery. Here we demonstrate that in low steroid conditions 1,25-dihydroxyvitamin D3 upregulates the expression of TRPV6, enhances the proliferation by increasing the number of cells entering into S-phase. We show that these pro-proliferative effects of 1,25-dihydroxyvitamin D3 are directly mediated via the overexpression of TRPV6 channel which increases calcium uptake into LNCaP cells. The apoptosis resistance of androgen-dependent LNCaP cells conferred by TRPV6 channel is drastically inversed when 1,25-dihydroxyvitamin D3 effects were combined with the successful TRPV6 knockdown. In addition, the use of androgen-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines allowed to suggest that the ability of 1,25-dihydroxyvitamin D3 to induce the expression of TRPV6 channel is a crucial determinant of the success or failure of 1,25-dihydroxyvitamin D3-based therapies.

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The effects of 1,25-dihydroxyvitamin D3 on androgen-independent cell lines.A, B, The effects of 1,25-dihydroxyvitamin D3 on androgen receptor-deficient DU-145 cell line in both 2 and 10% FCS-containing RPMI medium (A and B, respectively), * - P<0.05 (as compared to control), n = 3. C, D, The effects of 1,25-dihydroxyvitamin D3 on androgen-insensitive LNCaP C4-2 cell line in both 2 and 10% FCS-containing RPMI medium (C and D, respectively), * - P<0.05 (as compared to control), n = 3. E, the relative expression levels of TRPV6 channel in DU-145 cells treated with 100 µM 1,25-dihydroxyvitamin D3 for 3 days in 2 and 10% FCS-containing RPMI medium, * - P<0.05 (as compared to control), n = 3. F, the expression of TRPV6 channel induced by 100 nM 1,25-dihydroxyvitamin D3 for 3 days in LNCaP cells in steroid-deprived RPMI medium (LNCaP-ST), n = 3.
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pone-0016856-g004: The effects of 1,25-dihydroxyvitamin D3 on androgen-independent cell lines.A, B, The effects of 1,25-dihydroxyvitamin D3 on androgen receptor-deficient DU-145 cell line in both 2 and 10% FCS-containing RPMI medium (A and B, respectively), * - P<0.05 (as compared to control), n = 3. C, D, The effects of 1,25-dihydroxyvitamin D3 on androgen-insensitive LNCaP C4-2 cell line in both 2 and 10% FCS-containing RPMI medium (C and D, respectively), * - P<0.05 (as compared to control), n = 3. E, the relative expression levels of TRPV6 channel in DU-145 cells treated with 100 µM 1,25-dihydroxyvitamin D3 for 3 days in 2 and 10% FCS-containing RPMI medium, * - P<0.05 (as compared to control), n = 3. F, the expression of TRPV6 channel induced by 100 nM 1,25-dihydroxyvitamin D3 for 3 days in LNCaP cells in steroid-deprived RPMI medium (LNCaP-ST), n = 3.

Mentions: Two different androgen-independent cell lines were used: an androgen receptor-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines. Cells were cultivated in the same conditions of 2 or 10% FCS supplemented RPMI medium and the effects of 1,25-dihydroxyvitamin D3 were studied (Fig. 4). The effects of 1,25-dihydroxyvitamin D3 on androgen receptor deficient DU-145 cell line were likely to be serum-dependent since in 2% FCS the proproliferative effects of 1,25-dihydroxyvitamin D3 were conserved (Fig 4A), whereas in 10% FCS its effects were abolished (Fig. 4B). The other cell line insensitive to steroids, but still expressing the androgen receptor, LNCaP C4-2 was used, where the effects of 1,25-dihydroxyvitamin D3 were shown to be FCS-independent and 100 nM 1,25-dihydroxyvitamin D3 exerted its strong anti-proliferative effects (Fig. 4C–D). A real time quantitative PCR was performed showing the regulation of TRPV6 expression in DU-145 cells by 100 µM 1,25-dihydroxyvitamin D3 in both 2 and 10% FCS containing medium (Fig. 4E). Steroid-deprived conditions in the case of LNCaP cells (LNCaP-ST) were also used to confirm that the induction of TRPV6 expression strongly depends on the steroid content of the culture medium (Fig. 4F). Thus the pro-proliferative effects of 1,25-dihydroxyvitamin D3 on the growth of PCa cells are determined by its ability to induce the expression of TRPV6 channel and its induction appears to be strongly steroid-dependent.


TRPV6 determines the effect of vitamin D3 on prostate cancer cell growth.

Lehen'kyi V, Raphaël M, Oulidi A, Flourakis M, Khalimonchyk S, Kondratskyi A, Gordienko DV, Mauroy B, Bonnal JL, Skryma R, Prevarskaya N - PLoS ONE (2011)

The effects of 1,25-dihydroxyvitamin D3 on androgen-independent cell lines.A, B, The effects of 1,25-dihydroxyvitamin D3 on androgen receptor-deficient DU-145 cell line in both 2 and 10% FCS-containing RPMI medium (A and B, respectively), * - P<0.05 (as compared to control), n = 3. C, D, The effects of 1,25-dihydroxyvitamin D3 on androgen-insensitive LNCaP C4-2 cell line in both 2 and 10% FCS-containing RPMI medium (C and D, respectively), * - P<0.05 (as compared to control), n = 3. E, the relative expression levels of TRPV6 channel in DU-145 cells treated with 100 µM 1,25-dihydroxyvitamin D3 for 3 days in 2 and 10% FCS-containing RPMI medium, * - P<0.05 (as compared to control), n = 3. F, the expression of TRPV6 channel induced by 100 nM 1,25-dihydroxyvitamin D3 for 3 days in LNCaP cells in steroid-deprived RPMI medium (LNCaP-ST), n = 3.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3037935&req=5

pone-0016856-g004: The effects of 1,25-dihydroxyvitamin D3 on androgen-independent cell lines.A, B, The effects of 1,25-dihydroxyvitamin D3 on androgen receptor-deficient DU-145 cell line in both 2 and 10% FCS-containing RPMI medium (A and B, respectively), * - P<0.05 (as compared to control), n = 3. C, D, The effects of 1,25-dihydroxyvitamin D3 on androgen-insensitive LNCaP C4-2 cell line in both 2 and 10% FCS-containing RPMI medium (C and D, respectively), * - P<0.05 (as compared to control), n = 3. E, the relative expression levels of TRPV6 channel in DU-145 cells treated with 100 µM 1,25-dihydroxyvitamin D3 for 3 days in 2 and 10% FCS-containing RPMI medium, * - P<0.05 (as compared to control), n = 3. F, the expression of TRPV6 channel induced by 100 nM 1,25-dihydroxyvitamin D3 for 3 days in LNCaP cells in steroid-deprived RPMI medium (LNCaP-ST), n = 3.
Mentions: Two different androgen-independent cell lines were used: an androgen receptor-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines. Cells were cultivated in the same conditions of 2 or 10% FCS supplemented RPMI medium and the effects of 1,25-dihydroxyvitamin D3 were studied (Fig. 4). The effects of 1,25-dihydroxyvitamin D3 on androgen receptor deficient DU-145 cell line were likely to be serum-dependent since in 2% FCS the proproliferative effects of 1,25-dihydroxyvitamin D3 were conserved (Fig 4A), whereas in 10% FCS its effects were abolished (Fig. 4B). The other cell line insensitive to steroids, but still expressing the androgen receptor, LNCaP C4-2 was used, where the effects of 1,25-dihydroxyvitamin D3 were shown to be FCS-independent and 100 nM 1,25-dihydroxyvitamin D3 exerted its strong anti-proliferative effects (Fig. 4C–D). A real time quantitative PCR was performed showing the regulation of TRPV6 expression in DU-145 cells by 100 µM 1,25-dihydroxyvitamin D3 in both 2 and 10% FCS containing medium (Fig. 4E). Steroid-deprived conditions in the case of LNCaP cells (LNCaP-ST) were also used to confirm that the induction of TRPV6 expression strongly depends on the steroid content of the culture medium (Fig. 4F). Thus the pro-proliferative effects of 1,25-dihydroxyvitamin D3 on the growth of PCa cells are determined by its ability to induce the expression of TRPV6 channel and its induction appears to be strongly steroid-dependent.

Bottom Line: We show that these pro-proliferative effects of 1,25-dihydroxyvitamin D3 are directly mediated via the overexpression of TRPV6 channel which increases calcium uptake into LNCaP cells.The apoptosis resistance of androgen-dependent LNCaP cells conferred by TRPV6 channel is drastically inversed when 1,25-dihydroxyvitamin D3 effects were combined with the successful TRPV6 knockdown.In addition, the use of androgen-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines allowed to suggest that the ability of 1,25-dihydroxyvitamin D3 to induce the expression of TRPV6 channel is a crucial determinant of the success or failure of 1,25-dihydroxyvitamin D3-based therapies.

View Article: PubMed Central - PubMed

Affiliation: Inserm, U-1003, Equipe labellisée par la Ligue Nationale contre le cancer, Villeneuve d'Ascq, France.

ABSTRACT
Despite remarkable advances in the therapy and prevention of prostate cancer it is still the second cause of death from cancer in industrialized countries. Many therapies initially shown to be beneficial for the patients were abandoned due to the high drug resistance and the evolution rate of the tumors. One of the prospective therapeutical agents even used in the first stage clinical trials, 1,25-dihydroxyvitamin D3, was shown to be either unpredictable or inefficient in many cases. We have already shown that TRPV6 calcium channel, which is the direct target of 1,25-dihydroxyvitamin D3 receptor, positively controls prostate cancer proliferation and apoptosis resistance (Lehen'kyi et al., Oncogene, 2007). However, how the known 1,25-dihydroxyvitamin D3 antiproliferative effects may be compatible with the upregulation of pro-oncogenic TRPV6 channel remains a mystery. Here we demonstrate that in low steroid conditions 1,25-dihydroxyvitamin D3 upregulates the expression of TRPV6, enhances the proliferation by increasing the number of cells entering into S-phase. We show that these pro-proliferative effects of 1,25-dihydroxyvitamin D3 are directly mediated via the overexpression of TRPV6 channel which increases calcium uptake into LNCaP cells. The apoptosis resistance of androgen-dependent LNCaP cells conferred by TRPV6 channel is drastically inversed when 1,25-dihydroxyvitamin D3 effects were combined with the successful TRPV6 knockdown. In addition, the use of androgen-deficient DU-145 and androgen-insensitive LNCaP C4-2 cell lines allowed to suggest that the ability of 1,25-dihydroxyvitamin D3 to induce the expression of TRPV6 channel is a crucial determinant of the success or failure of 1,25-dihydroxyvitamin D3-based therapies.

Show MeSH
Related in: MedlinePlus