سید حمیدالله غفاری

Thyroid cancer is one of the most common endocrine malignancies. Anaplastic thyroid cancer is a rare and dead full cancer among types of the thyroid cancer. Despite the conventional chemotherapy, a considerable number of the patients show developing chemo resistance. Therefore, there is a necessary need to find the novel therapeutic approaches in the anaplastic thyroid cancer patients. The aim of this study was to study anti-tumor effect of Abemaciclib on the anaplastic thyroid carcinoma cell lines.

The human anaplastic thyroid cancer (SW1736 and C643) were cultured according to ATCC recommendations. The MTT assay was used to assess the chemo sensitivity of the cell lines in exposure to the desire concentration of Abemaciclib. Colony formation assay was used to determine the ability of the cell line colony formation in exposure to the drug. Quantitative real-time PCR was applied to analyze the mRNA expression of the apoptotic and anti-apoptotic genes.

The cell viability and proliferation of the thyroid cancer cell lines were remarkably inhibited in doses of 10 and 20 μM of Abemaciclib (p<0.0001). Also, Abemaciclib reduced the number of the SW1736 and C643 colonies in doses of 1 and 2.5 μM, respectively. Moreover, Abemaciclib significantly reduced anti-apoptotic gene expression levels, including BCL2 and CMYC, and increased pro-apoptotic gene expression levels, including p21 and BAX (p<0.05).

Our study suggests that Abemaciclib could be used as a therapeutic agent in anaplastic thyroid cancer. Further laboratory and animal studies are needed and recommended to evaluate the exact molecular and clinical properties of Abemaciclib.

Autophagy is a survival pathway required for cellular viability during starvation through catabolic self digestion of damaged proteins and organelles; however, autophagy may result in cell death if it proceeds to completion. Although the exact mechanism of this process is not clear, it seems that proper regulation of autophagy can potentially contribute to the therapeutics of cancers. The aim of this study was to determine the role of Arsenic trioxide (As2O3) in the induction of autophagy in human acute promyelocytic leukemia.

In this study, the role of autophagy in the As2O3-induced death of NB4 cells was assessed using electron microscopy. Also, quantitative real-time PCR method was used for expression of autophagy related genes in dose and Time dependent manner.

Our results showed that autophagy induces death in NB4 cells treated with arsenic trioxide. It was also found that the Atg7, Bclin1 and LC3 genes, which are essential genes for induction of autophagy, are the target of arsenic trioxide invasion.

This study showed As2O3-induced autophagy plays an important role in eliminating leukemia cells and treatment of cancer.

Since nearly 90% of patients with acute promyelocytic leukemia (APL) have high telomerase activity and significant shortened telomere length، these patients have، therefore، been suggested to be good candidates for the therapeutic intervention with telomerase inhibitors. This study was done to investigate the effects of BIBR1532، a non-nucleoside inhibitor of telomerase، on APL cells.

In this experimental study، for investigating the effect of BIBR1532، NB4 leukemic cells were cultured in the presence of various concentrations of BIBR1532. Succeeding apoptosis assay، Caspase-3 activity assay، and quantitative real-time PCR were applied to examine the effect of this drug on apoptosis percenage، enzymatic activity of Caspase-3، and quantitative expression of genes mRNA involved in apoptosis.

The results showed that BIBR1532 induced apoptosis in NB4 cells in a dose-dependent maner. Moreover، real time PCR results showed that BIBR1532 led to a significant decrease in mRNA of Bcl-2 gene and signficant increases in transcription of Bax، PUMA، and Caspase-3.

Since treatment with BIBR1532 could exert rapid apoptotic cell death in NB4 cells andactivate cellular apoptosis route، anti-telomerase-based therapy can regarded as a suitable strategy for APL treatment. Patients with progressive shortening of telomere length and high levels of telomerase activity are suitable candidates for treatment with telomerase inhibitors.

Tumor dissemination via blood to distant organs is the main cause of death. Therefore, there is a critical need to set up sensitive methods for the early detection of circulating tumor cells(CTCs) and disseminated tumor cells(DTCs) in peripheral blood (PB) and bone marrow(BM) specimens of breast cancer patients. The aim of this research is to study the detection of micrometastasis using MUC2 in such patients.

In this study, PB and BM samples were collected from 50 breast cancer patients after operation and before adjuvant therapy. Mucin 2 (MUC2) was used as a tumor marker and its transcript level in the sample patients was analyzed using gene specific, quantitative real-time PCR reaction with SYBR Green technology. Samples from 20 healthy individuals were used as negative controls. HPRT was used as a reference gene.

MUC2 mRNA was detected in 8 (16%) of PB and BM samples. MUC2 mRNA was not detected in PB samples of healthy individuals. The relapse rate among MUC2-positive patients was higher than MUC2-negative patients; and it was statistically significant in BM (P<0.05).

This study shows that MUC2 can be a suitable marker for the detection of micrometastasis in breast cancer patients at early stages of cancer and that it may provide the basis for identifying women at risk of relapse.

Acute promyelocytic leukemia (APL) is a distinct subtype of acute myeloid leukemia. APL is characterized by a balanced reciprocal translocation between chromosomes 15 and 17, t(5;17)). Important therapeutic strategies for this disease are ATRA and Arsenic trioxide. To eliminate tumor cells with arsenic, high dose of arsenic is needed. But high dose is toxic for normal tissue. The purpose of this study is Assessment of effect of low dose of arsenic trioxide in combination with AZT on NB4 cell line (APL cell line) to reduce toxic effect of high dose arsenic.

In this study after NB4 cell line culture and proliferation, the cells treated with low dose of arsenic trioxide(0.5µM) in combination with different doses of AZT(50, 100, 200 µM) and then viability and metabolic activity was assessed by try pan blue and MTT assay respectively.

Low dose of arsenic (0.5µm) alone and in combination with dose of 50µm of AZT has little effect on viability and metabolic activity but in combination with higher dose of AZT has significant effect on viability and metabolic activity and both viability and metabolic activity significantly reduced.Discussion and

Different apoptosis- induced mechanisms cause apoptosis by arsenic and AZT. Since some of these mechanisms between AZT and arsenic are similar, so maybe these similar mechanisms cause synergic effect and significant reduction of viability and metabolic activity in combination of these two drugs.

APL is a Prevalent leukemia that Approximately included 5-10% of patients with acute myeloblastic leukemia. ATRA and recently arsenic is used for treatment. ATRA leadsto resistance to treatment and arsenic is toxic in high doses.AZT induce cell death in different ways. The purpose of this study was Assessment of effect of AZT, a telomerase inhibitor, on NB4 cell line (APL cell line) to reduce toxic effect of high dose arsenic.

In this study, viability and metabolic activityof NB4 cells, treated by different concentrations of AZT(50,100,200 µM), was assessed by trypan blue dye method and MTT assay respectively.

Treated cells with AZT=50,100,200µM showed decreased viability, both in dose-dependent and time-dependent through trypan blue dye method and decreased cell metabolic activity by MTT assay.Discussion and

Considering that AZT is able to induce apoptosis and decrease cell activity, it seems AZT is a suitable drug for inhibiting the growth of tumor cells.

Transitional Cell Carcinoma (TCC) of bladder is the second most common urogenital malignancy and because of its high rate of recurrence (two third of tumors recur) vigilant surveillance is necessary. There have been a lot of efforts to find a proper biomarker for detecting urothelial cancers because available methods are expensive and invasive (like cystoscopy) or have a low degree of sensitivity. Urothelial malignancies, like other cancers tend to express a large amount of telomerase. The aim of this study was to evaluate the possible application of voided urine human telomerase reverse transcriptase (hTERT) mRNA assay in detecting low-grade bladder carcinoma in comparison with urine cytology

Chronic myeloid leukemia (CML) is a disorder of pluripotential hematopoietic stem cell that is as a myeloproliferative disease and occurs in about 15 percent of all leukemia. Two cell cycle regulatory proteins that function as tumor suppressor are P16INK4A and P14ARF. The origin of these two proteins is a human INK4A-ARF gene locus that located on chromosome 9p21. P16INK4A control retinoblastoma (Rb) and P14ARF control with p53 thought negative feedback. The purposes of this study, this was that whether these genes are preferable use as a factor in prognosis and progression of disease.

This research was a Cross sectional study. The expression of p16INK4A and p14ARF mRNA in about 73 peripheral bloods (PB) Samples were collected from 45 CML patients at different phases of disease were assayed by reverse transcriptase polymerase chain reaction (RT-PCR). 26 samples were from patients at chronic phase before any treatment, 26 samples 3 month after treatment with imatinib, 9 samples in accelerated phase and 12 samples in Blastic phase.Results. From 45 patients with CML, 33 patients (73%) were men and 12 patients (27%) were women. About 26 samples (35%) were p16INK4A positive and 55 samples (75%) were p14ARF RT-PCR positive. This expression of the two genes at different phases of disease were not statistically significant (p>0.05).

High percentage of the CML patients expressed P14ARF and P16INK4A genes. The expression of these gene at different phases of disease (diagnosis, accelerate, and Blastic phases) was not statistically significant; even though, the expression of these genes was higher after the treatment. The increased expression of these genes was probably because of the Imatinib treatment.