Chromatographic purification of proteins with cytotoxic potential from camel milk whey against cervical cancer cell line

Mahala, Neelam; Mittal, Aastha; Lal, Manohar; Dubey, Uma S.

doi:10.5958/2277-8934.2023.00007.3

Cited by 1 publication

(2 citation statements)

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“…Forty micrograms of protein lysate were loaded per well on a 12% SDS gel and run at a voltage of 80 V according to the procedure reported. [26] The separated protein was transferred onto the polyvinylidene difluoride (PVDF) membrane (Bio-Rad Laboratories). The membrane was blocked by 5% skimmed milk dissolved in tris buffer saline (TBS) at room temperature for 2 h.…”

Section: Western Blottingmentioning

confidence: 99%

“…The methodology is followed from our previously reported work. [26,27] Briefly, HEK 293 cells were harvested, counted, and seeded at 7000 cells per well in the 96-well plate for 24 h. The following day, cells were treated with various concentrations of the sorafenib-loaded casein nanoparticles and free sorafenib for 24 h. Untreated cells served as a negative control. Subsequently, 20 µL of 5 mg mL −1 MTT reagent was added to each well and incubated for 4 h. Lastly, the media was removed, and 150 µL of DMSO was added to the wells to solubilize the formazan crystals.…”

Section: Cell Viability Assaymentioning

confidence: 99%

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Evaluation of the cytotoxic activity of sorafenib‐loaded camel milk casein nanoparticles against hepatocarcinoma cells

Mittal,

Mahala,

Dhanawade

et al. 2024

Biotechnology Journal

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Sorafenib, a multikinase inhibitor is used to treat hepatocellular and renal carcinoma. However, a low solubility impedes its bioavailability and thus, effectiveness. This study aims to enhance its effectiveness by using novel camel milk casein nanoparticles as a delivery system. This study evaluates the cytotoxicity of sorafenib encapsulated in camel milk casein nanoparticles against human hepatocarcinoma cells (HepG2 cells) in vitro. Optimal drug loaded nanoparticles were stable for 1 month, had encapsulation efficiency of 96%, exhibited a particle size of 230 nm, zeta potential of −14.4 and poly disparity index of 0.261. Treatment with it led to cell morphology and DNA fragmentation as a characteristic of apoptosis. Flow cytometry showed G1 phase arrest of cell cycle and 26% increased apoptotic cells population upon treatment as compared to control. Sorafenib‐loaded casein nanoparticles showed 6‐fold increased ROS production in HepG2 cells as compared to 4‐fold increase shown by the free drug. Gene and protein expression studies done by qPCR and western blotting depicted upregulation of tumor suppressor gene p53, pro‐apoptotic Bax, and caspase‐3 along with downregulated anti‐apoptotic Bcl‐2 gene and protein expression which further emphasized death by apoptosis. It is concluded regarding the feasibility of these casein nanoparticles as a delivery system with enhanced therapeutic outcomes against hepatocellular carcinoma cells.

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Section: Western Blottingmentioning

confidence: 99%

Section: Cell Viability Assaymentioning

confidence: 99%