Aim To study the function of TRPC1/STIM1 in Ca²＋ entry and nitric oxide (NO) generation mediated by store-operated calcium channel (SOCC) and receptor-operated calcium channel (ROCC) in human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells were collected and cultured to the second~third passage. The constructed TRPC1 and STIM1 interference plasmids were transfected into human umbilical vein endothelial cells respectively, and the transfection efficiency was observed. The expressions of TRPC1 and STIM1 mRNA and protein were detected by real-time PCR and Western blot. The cells were incubated with CaR agonist spermine, CaR negative allosteric modulator Calhex231 and ROCC analogue TPA, protein kinase C (PKC) inhibitor Ro31-8220, PKCs and PKCμ inhibitor Go6967. Intracellular Ca²⁺ concentration ([Ca²⁺]_i) was detected using the fluorescence Ca²⁺ indicator Fura-2/AM, the production of NO was determined by DAF-FM of every group in HUVEC. The constructed TRPC1 and STIM1 interference plasmids were simultaneously transfected with human umbilical vein endothelial cells and incubated with CaR agonists to detect[Ca²＋]_i and NO production, and the interaction between STIM1 and TRPC1 was examined by co-immunoprecipitation. Results Compared with the control group, the expression of TRPC1 and STIM1 mRNA and protein in TRPC1 transfection group and STIM1 transfection group decreased obviously (P＜0.05). In four different treatment under the action of factors, the[Ca²＋]_i and the net NO fluorescence intensity ratio values of TRPC1 transfection group and STIM1 transfection group were significantly reduced (P＜0.05). Compared with control group, TRPC1 transfection group and STIM1 transfection group, the[Ca²＋]_i and the net NO fluorescence intensity ratio values of co-transfection group were significantly reduced (P＜0.05). TRPC1 and STIM1 interact to form a complex, and in the stimulation of CaR agonists under enhanced interaction. Conclusion TRPC1, STIM1 are components of SOCC and ROCC in Ca²＋ entry and NO generation in human umbilical vein endothelial cells.