Investigation of the effect of Lactarius deliciosus mushroom extract on CA-IX enzyme activity in melanoma cell lines

The carbonic anhydrase enzyme family (EC 4.2.1.1) are Zn2+-dependent metalloenzymes that produce HCO3- and H+ by catalyzing the hydration of CO2. The CA family of enzymes plays a key role in metabolism and tumorigenesis. CA-IX is a cell surface glycoprotein belonging to the CA family that regulates the pH of solid tumors. CA-IX is associated with cancer metastasis, progression, and response. Melanoma is a type of cancer with a high mortality rate resulting the from tumor formation of melanocyte cells that produce melanin in the epidermis. CA-IX is associated with the acidic microenvironment in melanoma. CA-IX The enzyme activity makes the tumor microenvironment more hypoxic. It has been reported that the expression of CA-IX is higher in melanoma cell lines in a hypoxic environment. In addition, antioxidant and anticancer effects of Lactarius deliciosus fungal species have been observed. The aim of this study is to determine the effect of a L. deliciosus mushroom extract in melanoma cell lines on the level of the enzyme CA-IX. In this study, an extract of L. deliciosus mushroom was prepared. Acetazolamide(AZA), a known inhibitor of CA-IX, was used for the control group. The cytotoxic effects of L. deliciosus mushroom extract and AZA in WM115 (primary) and SK-MEL-30 (metastatic) cell lines were evaluated by the WST-1 method, and IC50 values were calculated. The effects of AZA and the fungal extract on the enzyme levels of CA-IX were evaluated bythe Enzyme-Linked Immunosorbent Assay (ELISA). In the WM115 cell line the IC50 values for AZA and the mushroom extract of L. deliciosus were determined to be 21.41 μmol/L and 275 μg/ml, respectively. In the WM115 cells, the IC50 values determined to be 12.98 μmol/L and 276,45 μg/ml, respectively. Thus, AZA treatment had a significant inhibitory effect on the enzyme value CA-IX. It was found that L. deliciosus fungus extract statistically increased CA-IX enzyme value in melanoma cell lines SK-MEL-30 and WM115 (p<0.0001).