Curcumin Synergistically Enhances the Cytotoxicity of Arsenic Trioxide in U266 Cells by Increasing Arsenic Uptake

Despite the constant emergence of new methods for the treatment of multiple myeloma (MM), relapse and drug resistance still exist, especially in MM with p53 mutations. Arsenic trioxide (ATO) can be used in MM treatment, but this single drug has poor effectiveness and also side effects. Curcumin is a safe and effective compound that can enhance the anticancer effects of many drugs. Previous studies have suggested that tumor cell sensitivity to ATO is related to the intracellular arsenic content, and aquaporin 9 (AQP9) is the key factor that determines intracellular arsenic content. This study aimed to explore whether curcumin can increase ATO cytotoxicity in MM and whether the mechanism is related to the regulation of intracellular arsenic content. U266 was treated with ATO, curcumin, and their combination, and cell proliferation, apoptosis, and intracellular arsenic content were detected by CCK-8 assay, flow cytometry, and HPLC-ICP-MS, respectively. AQP9 mRNA and protein levels were detected by qPCR and western blotting. The levels of Mcl-1, Bcl-2, Bax, caspase-3, and cleaved caspase-3 protein were detected by western blotting. ATO-induced cytotoxicity to U266 occurred in a time- and dose-dependent manner, but the therapeutic efficacy at low drug concentrations was modest. The arsenic content in U266 was lower than that in NB4, and the arsenic uptake by U266 was concentration-dependent. The expression levels of AQP9 mRNA and AQP9 protein in U266 were lower than those in NB4. Curcumin significantly enhanced the lethality of ATO to U266. The arsenic content in U266 in the combined drug group increased significantly compared with ATO treatment alone. After curcumin treatment, the AQP9 mRNA and AQP9 protein expression levels in U266 also increased. Compared with the control group, the expression of antiapoptotic proteins Mcl-1 and Bcl-2 decreased, the expression of proapoptotic protein Bax increased, the ratio of Bax/Bcl-2 increased, and the expression of caspase-3 decreased and cleaved caspase-3 increased in the combined drug groups. Curcumin can enhance the killing effects of ATO on U266 by increasing the intracellular arsenic content, which may be related to the upregulation of AQP9 expression. The combination of these two drugs is expected to be a potential clinical treatment for MM.

Arsenic leads to autophagy of keratinocytes by increasing aquaporin 3 expression

Exposure to arsenic, a ubiquitous metalloid on Earth, results in human cancers. Skin cancer is the most common arsenical cancers. Both autophagy and aquaporin pathway are known to promote carcinogenesis. However, the mechanisms by which arsenic regulates aquaporin and autophagy in arsenical skin cancers remain elusive. This study aims to address how arsenic regulates aquaporin-3, the predominant aquaporin in epidermal keratinocytes, and how this process would induce autophagy. Quantitative real-time PCR and immunofluorescence were used to measure the expression of aquaporin 3 in arsenical skin cancers and arsenic-treated keratinocytes. Beclin-1 expression and autophagy were measured. We examined if blocking aquaporin 3 could interfere arsenic-induced autophagy in keratinocytes. Expression of aquaporin 3 is increased in arsenical cancers and in arsenic-treated keratinocytes. Arsenic induced autophagy in primary human keratinocytes. Notably, the arsenic-induced autophagy was inhibited by pretreatment of keratinocytes with aquaporin inhibitors Auphen or AgNO3, or RNA interference against aquaporin 3. The data indicates that the aquaporin 3 is an important cell membrane channel to mediate arsenic uptake and contributes to the arsenic-induced autophagy.

Determination of Arsenic Uptake Potential in An Edible Plant Species (Trigonellna foenum- granecum) and Assessment of Human Health Risk

Arsenic is a carcinogenic and toxic element that possesses a high health risk from its presence in crops, water, and soil. The present study has been conducted by fenugreek (Trigonella foenum-granecum) seeds which is a very common spice used for cooking, especially in India. An equal number of seeds have been germinated in laboratory conditions. Three concentrations e.g. 1, 2, and, 3 mg/L of arsenite (As+3) and arsenate (As+5) salt solutions were used throughout the experiments for the treatment of plants. After 10 days of germination, the concentration of the arsenic accumulated into the plant edible parts was estimated and health risk was assessed. Effects of arsenic concentration were observed through estimating the total chlorophyll (a, b, c), carotenoid content, and taking the fresh weight and dry weight of both the control and treatment plants. The results of the biochemical analysis revealed that chlorophyll and carotenoid contents were decreased than that of control plants. Moreover, fresh weight and dry weight results also showed lower values in treatments than in controls. The bioaccumulation factor results demonstrated that an increased level of soil arsenic doesn’t certainly result in high arsenic uptake by the Fenugreek plants. From the concentration estimated in the plant body, the health risk was assessed in adults and children and found that both adults and children having a potential health risk upon consumption of fenugreek. Moreover, Incremental Life Time Cancer Risk was found high (< 10-4) which indicates the presence of potential cancer risk. Hence, it is possible to conclude from the present study that fenugreek can bio-accumulate arsenic and it may be used as an indicator plant for arsenic-contaminated areas.