Apoptosis induction in colon cancer cells (SW480) by BiFe2O4@Ag nanocomposite synthesized from Chlorella vulgaris extract and evaluation the expression of CASP8, BAX and BCL2 genes.

BACKGROUND: The use of nanomaterials in cancer diagnosis and treatment has received considerable interest. Preparation of nanoscale complex molecules could be considered to improve the efficacy and minimize toxicity of the product. This work aimed to biosynthesize BiFe2O4@Ag nanocomposite using the Chlorella vulgaris extract and its cytotoxic effect on colon cancer cell line. METHODS: The physicochemical properties of the bioengineered BiFe2O4 @Ag were investigated by Transmission Electron Microscopy (TEM), Field Emission Scanning Electron Microscopy (FE-SEM), Zeta potential, Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FT-IR), Energy Dispersive X-ray Spectroscopy (EDX), Vibrating-sample Magnetometer (VSM) and X-ray Diffraction Analysis (XRD). The cytotoxic potential of BiFe2O4 @Ag was evaluated by MTT assay against SW480 colon cancer cell line. The expression levels of apoptotic genes including BAX, BCL2 and CASP8 were determined by Real-time PCR. The rate of apoptosis and necrosis of the cancer cells as well as the cell cycle analysis were evaluated by flow cytometry. RESULTS: Physicochemical assays indicated the nanoscale synthesis (10-70 nm) and functionalization of BiFe2O4 nanoparticles by Ag atoms. The VSM analysis revealed the magnetism of BiFe2O4 @Ag nanocomposite. According to the MTT assay, colon cancer cells (SW480) were considerably more sensitive to BiFe2O4 @Ag nanocomposite than normal cells. Apoptotic cell percentage increased from 1.93% to 73.66%, after exposure to the nanocomposite. Cell cycle analysis confirmed an increase in the number of the cells in subG1 and G0/G1 phases among nanocomposite treated cells. Moreover, treating the colon cancer cells with BiFe2O4 @Ag caused an increase in the expression of CASP8, BAX, and BCL2 genes by 3.1, 2.6, and 1.2 folds, respectively. Moreover, activity of Caspase-3 protein increased by 2.4 folds and apoptotic morphological changes appeared which confirms that exposure to the nanocomposite induces extrinsic pathway of apoptosis in colon cancer cells. CONCLUSION: The considerable anticancer potential of the synthesized BiFe2O4 @Ag nanocomposite seems to be related to the induction of oxidative stress which leads to inhibit cell cycle progression and cell proliferation. This study reveals that the BiFe2O4 @Ag is a potent compound to be used in biomedical fields.

Synthesize of Bi2O3/Gln-TSC nanoparticles and evaluation of their toxicity on prostate cancer cells and expression of CASP8, BAX, and Bcl-2 genes.

Due to the high prevalence and considerable increase of prostate cancer, finding novel therapeutic compounds for the treatment of prostatic cancer has been the goal of many researches. In this study, we aimed to fabricate the Bismuth oxide (Bi2O3) NPs, functionalized with glutamine (Gln) and conjugated with Thiosemicarbazide (TSC). Then, the anticancer mechanism of the synthesized NPs was investigated using the cellular and molecular tests including MTT assay, Flow cytometry, Caspase-3 activity assay, Hoechst staining and Real Time PCR. The FT-IR and XRD assays confirmed the identity of the synthesized Bi2O3/Gln-TSC NPs. The size range of the synthesized spherical particles was 10-60 nm and the zeta potential was - 23.8 mV. The purity of the NPs was confirmed by EDX-mapping analysis. The Bi2O3/Gln-TSC was considerably more toxic for prostate cancer cells than normal human cells and the IC50 was calculated 35.4 and 305 µg/mL, respectively. The exposure to the NPs significantly increased the frequency of apoptotic cells from 4.7 to 75.3%. Moreover, the expression of the CASP8, BAX, and Bcl-2 genes after exposure to the NPs increased by 2.8, 2.3, and 1.39 folds. Treating the cancer cells with Bi2O3/Gln-TSC increased the activity of the Caspase-3 protein and apoptotic morphological features were observed by Hoechst staining in the treated cells. This work showed that Bi2O3/Gln-TSC has considerable cytotoxicity for prostate cancer cells and could inducing both intrinsic and extrinsic pathways of apoptosis.

Comparison of high-resolution melting analysis with direct sequencing for detection of FLT3-TKD, FLT3-ITD and WT1 mutations in acute myeloid leukemia.

BACKGROUND: Acute Myeloid Leukemia (AML) is a group of hematologic diseases characterized by a variety of clinically important genetic alterations. Genetic mutations affecting the FMS-like receptor tyrosine kinase-3 (FLT3) and Wilm's tumor (WT-1) genes are associated with poor prognosis in AML. In this work, efficiency of HRM method for detection of FLT3-ITD, FLT3-TKD, and WT-1 mutations was assessed in comparison with direct sequencing. METHOD: A total of 58 formalin-fixed, paraffin-embedded BM biopsy specimens of AML patients were analyzed. Mutation detection was performed by HRM method and the results were consequently compared with direct sequencing RESULTS: FLT3 and WT-1 mutations were detected in 21 (36.2%) and 3 (5.17%) samples, respectively. Among all FLT3 mutations, 10 (17.2%) and 11 (18.2%) samples were harboring the FLT3-ITD and-TKD gene mutations, respectively. Frequency of the FLT3-ITD was not statistically different in females (51%) and males (49%). Also, FLT3-TKD was more common in males although the differences in gender distribution were not statistically significant (P = 0.721 and P = 0.626, respectively). CONCLUSIONS: Regarded as the desirable characteristic, the present study is generally distinguished by the similar previous ones due to assessing the FFPE BM tissue from the perspective of the type of assessed sample. This discrepancy between our results and those in prior studies may be due to the disparity of the studied population size, adopted methods as well as the sample type. In this survey, regarding to low amount of extracted DNA from the paraffinized samples, the HRM method was efficient in determining the mentioned mutations.