Hepatocellular carcinoma (HCC) is a lethal complication in the human liver. The most important problems with HCC are that the tumor appears in several areas of the liver, it is difficult to diagnose and identify, and there is a possibility of tumor recurrence (1). Therefore, identification of diagnostic biomarkers of HCC and effective therapeutic agents is very necessary. Organic cation transporters are integral transmembrane factors with critical effects in metabolism and detoxification via uptake of endogenous and exogenous elements and anticancer drugs. Organic cation transporter 3 (OCT3), as a transporting protein in human, can remove small organic cations created by environmental toxins and/or drugs (2)....

Organic cation transporter 3 (OCT3) as a high-capacity transporter contribute to the metabolism of metformin. The present study was conducted to determine the genotype frequencies of the variant OCT3-1233G>A (rs2292334) in patients with newly diagnosed type 2 diabetes (T2D) and its relationship with response to metformin. Materials and Methods: This study included 150 patients with T2D who were classified into two groups following three months of metformin therapy: responders (by more than 1% reduction in HbA1c from baseline) and nonresponders (less than 1% reduction in HbA1c from baseline). PCR-based restriction fragment length polymorphism (RFLP) served to genotype OCT3-564G>A variant. Results: The parameters such as HbA1c (P<0. 001) and BMI (P<0. 001) in both patients with GA + AA genotype and GG genotype decreased significantly following 3 months of metformin therapy compared with baseline. The mean reduction in HbA1c levels following 3 months was higher in patients with the A allele (0. 77% reduction from baseline) than in those with the homozygous G allele (0. 54% reduction from baseline). Also, in GA + AA genotypes compared with GG genotypes, the mean reduction in HbA1c values from baseline was 0. 34% for responders and 0. 14% for non-responders. Conclusion: Considering the roles of genetic variations in the function of metformin transporters, the effect of variations such as 1233G>A in the OCT3, which is a high-capacity transporter widely expressed in various tissues cannot be ignored. Comparing the allele frequencies of OCT3-1233G>A variant in our study and different ethnic populations confirm that the variant is a highly polymorphic variant.

Background: Organic cation transporter 3 (OCT3) is an excellent transporter for metformin, which is used as first-line therapy for type 2 diabetes (T2D). OCT3 genetic variants may influence the clinical response to metformin. This study aimed to determine the genotype and allele frequency of OCT3-564G>A (rs3088442) variant and its role in the glycemic response to metformin in patients with newly diagnosed T2D. Materials and Methods: Based on the response to metformin, 150 patients were classified into two groups: Sixty-nine responders (decrease in glycated hemoglobin [HbA1c] values by more than 1% from the baseline) and 81 nonresponders (decrease in HbA1c values<1% from the baseline). HbA1c levels were determined by chromatography. The variant OCT3-564G>A was genotyped using polymerase chain reaction - based restriction fragment length polymorphism.Results: The genotypes frequencies were 51.3% GG, 36% AG, and 12.7% AA. Allele frequency of major allele (G) and minor allele (A) in OCT3-564G>A variant was found to be 0.69 and 0.31, respectively. Fasting glucose, HbA1c, body mass index, and lipid profile in both GG genotypes and GA+AA group decreased significantly after 3 months of metformin therapy compared with baseline (P<0.05). In both responders and nonresponders, HbA1c and fasting glucose levels were lower in patients with the GA+AA genotype than in those with the GG genotype; however, the differences were not statistically significant (P>0.05).Conclusion: The A allele frequency (which may be a protective allele against coronary heart disease) in the Iranian diabetic patients was lower than Iranian, Caucasian and Japanese healthy populations. Metformin is useful in improving the lipid profile, in addition to its impacts in glycemic control, and these effects are regardless of OCT3-564G>A variant.

Objective: Generation of patient specific stem cell is among the ultimate goals in regenerative medicine. On the other hand contact between matrix proteins and integrin receptors adjust many cell’s function such as migration during embryogenesis, immune responses and tumour invasion. In this study we de-differentiated NIH3T3 cells by mouse Embryonic Stem Cell (mESC) extract in order to determine the expression level of pluripotency markers as well as a2b1 and a5b1 integrins.Materials and Methods: Mouse embryonic stem cell extracts were prepared and NIH3T3 cells were reprogrammed by mESC extract. Generated iPS (induced Pluripotent Stem cells) colonies were picked up in the sterile conditions and checked for the pluripotency by alkaline phosphatase kit, Reverse Transcriptase PCR (RT-PCR) for oct3/4 and immunocytochemistry for oct3/4, sox2 and nanog. Then the expression level of oct3/4, a2b1 and a5b1 integrins was studied by RTPCR.Results: NIH3T3 cells treated with mESC extracts showed noticeable changes in cell morphology as early as day 3 post-treatment forming colonies similar to typical mESC morphology by day 10, after three passages. Alkaline phosphatase test and immunocytochemistry staining were performed for the iPS colonies. The results indicated that these colonies not only showed the alkaline phosphatase activity, but also the expression of sox2, oct3/4 and nanog by their specific antibodies in immunocytochemistry. Also RT-PCR results revealed the expression of oct3/4 a2b1 and a5b1 integrins in iPS cells.Conclusion: These data provide evidence for the generation of iPS cells from differentiated somatic cells by mESC extract also the expression of a2b1 and a5b1 integrins in iPS cells.

Background: Cancer stem cells (CSCs) substantially influence the development of colorectal cancer (CRC), metastasis, relapse, and resistance to therapy. Ibuprofen and hyperthermia can be effective in the treatment of cancer. Herein, we evaluated the effects of hyperthermia and ibuprofen on the isolated-CSCs of CRC. Methods: This experimental study was conducted between Sep 2020 and Jan 2022 at the Department of Pathology, School of Medicine, Shiraz University of Medical Sciences, Iran. A non-adhesive culture system was used to isolate CSCs from HT-29 cells. To confirm the stemness nature of isolated-CSCs, the expression of stemness genes and protein markers was evaluated by quantitative Real-time PCR (qRT-PCR) and flow cytometry assay. The isolated-CSCs were treated with hyperthermia and ibuprofen. The cell viability was determined by MTT assay and trypan blue staining. The expression of stemness, proliferation, Wnt signaling pathway and apoptosis genes was assessed by qRT-PCR. Results: CSCs were isolated within 14 days. The expression of CD-133 marker and OCT3/4, C-MYC, KLF4, and NANOG genes in isolated-CSCs was higher than HT-29 cells (P<0.05). Cell viability of treated-CSCs were considerably reduced (P<0.05). Hperthermia reduced the expression of OCT3/4, NANOG, PCNA, WNT1 and CTNNB1 genes and increased the expression of P53, BAX, and KLF4 genes (P<0.05). Ibuprofen decreased the expression of OCT3/4, BCL2, NANOG, PCNA, WNT1, and CTNNB1 genes and increased the expression of P53, BAX, and KLF4 genes in treated-CSCs (P<0.05). Conclusion: Hyperthermia and ibuprofen treatment demonstrate an inhibitory effect on colorectal CSCs. However, using combination therapy is remaining to be tested.

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of blastocyst and differentiate into all three embryonic germ layers: ectoderm, endoderm, and mesoderm. In this study, ESCs are derived from Hand Made Cloning (HMG) blastocysts and their efficiencies compared to ESCs derived from In Vitro Fertilization (IVF) embryos. Feeder layer was used for ESCs culture, and culture medium consisting of Knockout- Dulbecco’s Modified Eagle’s Medium (Ko-DMEM) supplemented with Knockout Serum Replacement (KSR), Leukemia Inhibitory Factor (LIF), Basic Fibroblast Growth Factor-2 (FGF-2), L-glutamine, nonessential amino acids and gentamicin. The cell surface antigens used for characterization were the SSEA-1, SSEA-4, TRA-1-60 and TRA-1-81 and the pluripotency markers were NANOG, OCT3/4 and SOX2. Results showed that, the growth rate of ESCs colonies in ESCs from IVF embryos was significantly higher than ESCs from HMG embryos (120% compared with 65%, respectively). Not only real-time PCR results revealed the same expression level of SOX2, OCT3/4 and cMYC between them, but also ESCs from HMG embryos resulted to higher expression of NANOG. Both of ESCs groups maintain in pluripotency state for more than two years and differentiated to the different types of cells like neuron, epithelial, lipid and muscle cells.

Purpose: Previous studies have documented that cumulus granulosa cells (GCs) can transdifferentiation into different non-ovarian cells, showing their multipotentiality to repopulate the injured cells in ovarian tissue. The current experiment is aimed to assess the differentiation capacity of human cumulus GCs toward the oocyte-like phenotype in vitro. Methods: GCs were isolated from healthy female volunteers subjected to in vitro fertilization or intra-cytoplasmic sperm injection (IVF-ICSI). The effect of different media supplemented with leukemia inhibitory factors (LIFs), 5 ng/mL estradiol, and 0. 005 IU/mL follicle-stimulating hormone (FSH) were investigated to the differentiation of GCs toward oocyte-like phenotype via monitoring the expression of Oct3/4 and GATA-4 using flow cytometry analysis. The expression of genes such as FIGLA, NOBOX, and SYCP3 was measured by real-time polymerase chain reaction (PCR) assay. We also assess morphological adaptation by using bright-field microscopic imaging. Results: Exposure of GCs to LIFs increased the number of cells expressing stemness factor Oct3/4 coincided with the suppression of GATA-4 after 7 days (P < 0. 05). We found that the transcript level of all genes FIGLA, Nobox, and SYCP-3 decreased in cells after treatment with a FSH (P < 0. 05). According to our data, the incubation of GCs with estradiol increased the expression of genes related to the oocyte-like phenotype. Conclusion: Our finding revealed that the combination of LIFs and estradiol could induce the GCs oogenesis capacity and thereby is possibly suggested as a therapeutic strategy during the occurrence of gynecological disorders.