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T-type calcium channel antagonists, mibefradil and NNC-55-0396 inhibit cell proliferation and induce cell apoptosis in leukemia cell lines.

Huang W, Lu C, Wu Y, Ouyang S, Chen Y - J. Exp. Clin. Cancer Res. (2015)

Bottom Line: T-type Ca(2+) channels are often aberrantly expressed in different human cancers and participate in the regulation of cell cycle progression, proliferation and death.Mechanistically, these inhibitors played a dual role on cell viability: (i) blunting proliferation, through a halt in the progression to the G1-S phase; and (ii) promoting cell apoptosis, partially dependent on the endoplasmic reticulum Ca(2+) release.In addition, we observed a reduced phosphorylation of ERK1/2 in MOLT-4 cells in response to mibefradil and NNC-55-0396 treatment.

View Article: PubMed Central - PubMed

Affiliation: Fujian Institute of Hematology, Fujian Medical University Union Hospital, Fuzhou, 350004, People's Republic of China. hwf0625@163.com.

ABSTRACT

Background: T-type Ca(2+) channels are often aberrantly expressed in different human cancers and participate in the regulation of cell cycle progression, proliferation and death.

Methods: RT-PCR, Q-PCR, western blotting and whole-cell patch-clamp recording were employed to assess the expression of T-type Ca(2+) channels in leukemia cell lines. The function of T-type Ca(2+) channels in leukemia cell growth and the possible mechanism of the effect of T-type Ca(2+) channel antagonists on cell proliferation and apoptosis were examined in T-lymphoma cell lines.

Results: We show that leukemia cell lines exhibited reduced cell growth when treated with T-type Ca(2+) channel inhibitors, mibefradil and NNC-55-0396 in a concentration-dependent manner. Mechanistically, these inhibitors played a dual role on cell viability: (i) blunting proliferation, through a halt in the progression to the G1-S phase; and (ii) promoting cell apoptosis, partially dependent on the endoplasmic reticulum Ca(2+) release. In addition, we observed a reduced phosphorylation of ERK1/2 in MOLT-4 cells in response to mibefradil and NNC-55-0396 treatment.

Conclusions: These results indicate that mibefradil and NNC-55-0396 regulate proliferation and apoptosis in T-type Ca(2+) channel expressing leukemia cell lines and suggest a potential therapeutic target for leukemia.

No MeSH data available.


Related in: MedlinePlus

Effect of T-type Ca2+ channel blockers, mibefradil and NNC-55-0396 on cell growth. MOLT-4 (a), Jurkat (b), U937 (c) and HEL (d) cells were cultured in the present of mibefradil or NNC-55-0396 (2–10 μM) for 48 h. All data points represent an average of three to five experiments (± SEM). *P < 0.05 versus control, **P < 0.01 versus control
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Fig2: Effect of T-type Ca2+ channel blockers, mibefradil and NNC-55-0396 on cell growth. MOLT-4 (a), Jurkat (b), U937 (c) and HEL (d) cells were cultured in the present of mibefradil or NNC-55-0396 (2–10 μM) for 48 h. All data points represent an average of three to five experiments (± SEM). *P < 0.05 versus control, **P < 0.01 versus control

Mentions: To evaluate the functional expression of T-type Ca2+ channels in MOLT-4 T cells, whole-cell patch-clamp recordings were performed to record the T-type Ca2+ current. Using Ba2+ as a charge carrier, the current activated at −30 mV, with peak current at 0 mV, and displayed rapid activation and inactivation kinetics (Additional file 1: Figure S1). The amplitude of T-current in MOLT-4 cells varied between 10 and 20 pA (n = 8), and the mean T-current density was 0.69 ± 0.15 pA/pF. Figure 1c shows that a ~15 pA T-type Ca2+ current was elicited by a depolarizing pulse at 0 mV when held at −80 mV (control). After perfusion of 2 μM of NNC-55-0396, the T-type Ca2+ current was inhibited by ~70 %. In addition, MOLT-4 cells showed a mean resting potential of −30.5 ± 1.8 mV (n = 12) and membrane capacitance of 14.5 ± 0.7 pF (n = 15). Treatment of T lymphocytes with mibefradil, a selective inhibitor against T-type Ca2+ channels, blocked Ca2+ influx (Additional file 2: Figure S2). These findings indicate that T-type Ca2+ channels play a significant role in the Ca2+ influx pathways of human leukemia T cell line.Fig. 2


T-type calcium channel antagonists, mibefradil and NNC-55-0396 inhibit cell proliferation and induce cell apoptosis in leukemia cell lines.

Huang W, Lu C, Wu Y, Ouyang S, Chen Y - J. Exp. Clin. Cancer Res. (2015)

Effect of T-type Ca2+ channel blockers, mibefradil and NNC-55-0396 on cell growth. MOLT-4 (a), Jurkat (b), U937 (c) and HEL (d) cells were cultured in the present of mibefradil or NNC-55-0396 (2–10 μM) for 48 h. All data points represent an average of three to five experiments (± SEM). *P < 0.05 versus control, **P < 0.01 versus control
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4443536&req=5

Fig2: Effect of T-type Ca2+ channel blockers, mibefradil and NNC-55-0396 on cell growth. MOLT-4 (a), Jurkat (b), U937 (c) and HEL (d) cells were cultured in the present of mibefradil or NNC-55-0396 (2–10 μM) for 48 h. All data points represent an average of three to five experiments (± SEM). *P < 0.05 versus control, **P < 0.01 versus control
Mentions: To evaluate the functional expression of T-type Ca2+ channels in MOLT-4 T cells, whole-cell patch-clamp recordings were performed to record the T-type Ca2+ current. Using Ba2+ as a charge carrier, the current activated at −30 mV, with peak current at 0 mV, and displayed rapid activation and inactivation kinetics (Additional file 1: Figure S1). The amplitude of T-current in MOLT-4 cells varied between 10 and 20 pA (n = 8), and the mean T-current density was 0.69 ± 0.15 pA/pF. Figure 1c shows that a ~15 pA T-type Ca2+ current was elicited by a depolarizing pulse at 0 mV when held at −80 mV (control). After perfusion of 2 μM of NNC-55-0396, the T-type Ca2+ current was inhibited by ~70 %. In addition, MOLT-4 cells showed a mean resting potential of −30.5 ± 1.8 mV (n = 12) and membrane capacitance of 14.5 ± 0.7 pF (n = 15). Treatment of T lymphocytes with mibefradil, a selective inhibitor against T-type Ca2+ channels, blocked Ca2+ influx (Additional file 2: Figure S2). These findings indicate that T-type Ca2+ channels play a significant role in the Ca2+ influx pathways of human leukemia T cell line.Fig. 2

Bottom Line: T-type Ca(2+) channels are often aberrantly expressed in different human cancers and participate in the regulation of cell cycle progression, proliferation and death.Mechanistically, these inhibitors played a dual role on cell viability: (i) blunting proliferation, through a halt in the progression to the G1-S phase; and (ii) promoting cell apoptosis, partially dependent on the endoplasmic reticulum Ca(2+) release.In addition, we observed a reduced phosphorylation of ERK1/2 in MOLT-4 cells in response to mibefradil and NNC-55-0396 treatment.

View Article: PubMed Central - PubMed

Affiliation: Fujian Institute of Hematology, Fujian Medical University Union Hospital, Fuzhou, 350004, People's Republic of China. hwf0625@163.com.

ABSTRACT

Background: T-type Ca(2+) channels are often aberrantly expressed in different human cancers and participate in the regulation of cell cycle progression, proliferation and death.

Methods: RT-PCR, Q-PCR, western blotting and whole-cell patch-clamp recording were employed to assess the expression of T-type Ca(2+) channels in leukemia cell lines. The function of T-type Ca(2+) channels in leukemia cell growth and the possible mechanism of the effect of T-type Ca(2+) channel antagonists on cell proliferation and apoptosis were examined in T-lymphoma cell lines.

Results: We show that leukemia cell lines exhibited reduced cell growth when treated with T-type Ca(2+) channel inhibitors, mibefradil and NNC-55-0396 in a concentration-dependent manner. Mechanistically, these inhibitors played a dual role on cell viability: (i) blunting proliferation, through a halt in the progression to the G1-S phase; and (ii) promoting cell apoptosis, partially dependent on the endoplasmic reticulum Ca(2+) release. In addition, we observed a reduced phosphorylation of ERK1/2 in MOLT-4 cells in response to mibefradil and NNC-55-0396 treatment.

Conclusions: These results indicate that mibefradil and NNC-55-0396 regulate proliferation and apoptosis in T-type Ca(2+) channel expressing leukemia cell lines and suggest a potential therapeutic target for leukemia.

No MeSH data available.


Related in: MedlinePlus