Exogenous administration of hydrogen sulfide (H₂S) is emerging as an alternative anticancer treatment. H₂S-releasing compounds have been shown to exert a strong anticancer effect by suppressing proliferation and/or inducing apoptosis in several cancer cell types, including colorectal carcinoma (CRC). The mechanism whereby exogenous H₂S affects CRC cell proliferation is yet to be clearly elucidated, but it could involve an increase in intracellular Ca²⁺ concentration ([Ca²⁺]_i). Herein, we sought to assess for the first time whether (and how) sodium hydrosulfide (NaHS), one of the most widely employed H₂S donors, induced intracellular Ca²⁺ signals in primary cultures of human metastatic CRC (mCRC) cells. We provided the evidence that NaHS induced extracellular Ca²⁺ entry in mCRC cells by activating the Ca²⁺-permeable channel Transient Receptor Potential Vanilloid 1 (TRPV1) followed by the Na⁺-dependent recruitment of the reverse-mode of the Na⁺/Ca²⁺ (NCX) exchanger. In agreement with these observations, TRPV1 protein was expressed and capsaicin, a selective TRPV1 agonist, induced Ca²⁺ influx by engaging both TRPV1 and NCX in mCRC cells. Finally, NaHS reduced mCRC cell proliferation, but did not promote apoptosis or aberrant mitochondrial depolarization. These data support the notion that exogenous administration of H₂S may prevent mCRC cell proliferation through an increase in [Ca²⁺]_i, which is triggered by TRPV1.

(1) Background: Store-operated Ca²⁺ entry (SOCE) drives the cytotoxic activity of cytotoxic T lymphocytes (CTLs) against cancer cells. However, SOCE can be enhanced in cancer cells due to an increase in the expression and/or function of its underlying molecular components, i.e., STIM1 and Orai1. Herein, we evaluated the SOCE expression and function in tumour-infiltrating lymphocytes (TILs) from metastatic colorectal cancer (mCRC) patients. (2) Methods: Functional studies were conducted in TILs expanded ex vivo from CRC liver metastases. Peripheral blood T cells from healthy donors (hPBTs) and mCRC patients (cPBTs) were used as controls. (3) Results: SOCE amplitude is enhanced in TILs compared to hPBTs and cPBTs, but the STIM1 protein is only up-regulated in TILs. Pharmacological manipulation showed that the increase in SOCE mainly depends on tonic modulation by diacylglycerol kinase, which prevents the protein kinase C-dependent inhibition of SOCE activity. The larger SOCE caused a stronger Ca²⁺ response to T-cell receptor stimulation by autologous mCRC cells. Reducing Ca²⁺ influx with BTP-2 during target cell killing significantly increases cytotoxic activity at low target:effector ratios. (4) Conclusions: SOCE is enhanced in ex vivo-expanded TILs deriving from mCRC patients but decreasing Ca²⁺ influx with BTP-2 increases cytotoxic activity at a low TIL density.