Measuring Ca~(2+) influxes of TRPC1-dependent Ca~(2+) channels in HL-7702 cells with Non-invasive Mi

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:shuwenglei
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AIM:To explore the possibility of using the Non-invasive Micro-test Technique(NMT)to investigate the role of Transient Receptor Potential Canonical 1(TRPC1) in regulating Ca 2+ influxes in HL-7702 cells,a normal human liver cell line.METHODS:Net Ca 2+ fluxes were measured with NMT, a technology that can obtain dynamic information of specific/selective ionic/molecular activities on material surfaces,non-invasively.The expression levels of TRPC1 were increased by liposomal transfection,whose effectiveness was evaluated by Western-blotting and single cell reverse transcription-polymerase chain reaction. RESULTS:Ca 2+ influxes could be elicited by adding 1 mmol/L CaCl2 to the test solution of HL-7702 cells.They were enhanced by addition of 20μmol/L noradrenalin and inhibited by 100μmol/L LaCl3(a non-selective Ca 2+ channel blocker);5μmol/L nifedipine did not induce any change.Overexpression of TRPC1 caused increased Ca 2+ influx.Five micromoles per liter nifedipine did not inhibit this elevation,whereas 100μmol/L LaCl3 did.CONCLUSION:In HL-7702 cells,there is a type of TRPC1-dependent Ca 2+ channel,which could be detected via NMT and inhibited by La3+. AIM: To explore the possibility of using the Non-invasive Micro-test Technique (NMT) to investigate the role of Transient Receptor Potential Canonical 1 (TRPC1) in regulating Ca 2+ influxes in HL-7702 cells, a normal human liver cell line .METHODS: Net Ca 2+ fluxes were measured with NMT, a technology that can obtain dynamic information of specific / selective ionic / molecular activities on material surfaces, non-invasively. The expression levels of TRPC1 were increased by liposomal transfection, whose effectiveness was evaluated by Western-blotting and single cell reverse transcription-polymerase chain reaction. RESULTS: Ca 2+ influxes could be elicited by adding 1 mmol / L CaCl 2 to the test solution of HL-7702 cells. They were enhanced by addition of 20 μmol / L noradrenalin and inhibited by 100 μmol / L LaCl3 (a non-selective Ca 2+ channel blocker); 5 μmol / L nifedipine did not induce any change. Overexpression of TRPC1 caused increased Ca 2+ influx. Failed micromoles per liter nifedipine did not inhibit this elevation, and 100 μmol / L LaCl3 did. CONCLUSION: In HL-7702 cells, there is a type of TRPC1-dependent Ca 2+ channel, which could be detected via NMT and inhibited by La3 +.
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