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Measuring Ca2+ influxes of TRPC1-dependent Ca2+ channels in HL-7702 cells with non-invasive micro-test technique
Zhen-Ya Zhang1, Wen-Jun Wang, Li-Jie Pan
1Department of General Surgery, Digestive Medical Center, the First Affiliated Hospital, Medical School, Tsinghua University, Beijing 100016, China.
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 micromol/L noradrenaline and inhibited by 100 micromol/L LaCl(3) (a non-selective Ca(2+) channel blocker); 5 micromol/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 100 micromol/L LaCl(3) 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 La(3+).
