The effects of chemotherapeutic drugs on PD-L1 gene expression in breast cancer cell lines.

Breast cancer is the most common cancer among women in terms of prevalence and mortality, and chemotherapy is one of the most effective treatments at higher stages. However, resistance to chemotherapy is the main obstacle in the treatment of this cancer. Accumulated evidence identified the PD-L1 protein as an essential protein in the development of different cancers. Abnormal expression of this protein in various tumor cells is linked to cancer development and inhibiting the function of immune cells, which correlated with reduced beneficial effects of chemotherapy drugs. In the present study, the effects of common chemotherapy drugs including doxorubicin, paclitaxel, and docetaxel on the expression of the PD-L1 gene were investigated by qRT-PCR before and after the treatment with these drugs in MD231, MD468, SKBR3 breast cancer cell lines. Also, the MTT test was applied to examine the effects of drugs on the growth and proliferation of cancer cells considering PD-L1 expression. The expression of the PD-L1 gene increased after 24 and 48 h of treatment with chemotherapy drugs. The obtained results indicate the enhancing effects of chemotherapy drugs on PD-L1 gene expression, which have a suppressive effect on the immune system against breast cancer. The use of these drugs as the first line of chemotherapy in triple-negative breast cancer is not recommended. However, there is still a need for further experimental and clinical research on the exact effects of these drugs on undesired immune cells exhaustion in breast cancer therapy.

Silencing ZEB2 Induces Apoptosis and Reduces Viability in Glioblastoma Cell Lines.

BACKGROUND: Glioma is an aggressive type of brain tumor that originated from neuroglia cells, accounts for about 80% of all malignant brain tumors. Glioma aggressiveness has been associated with extreme cell proliferation, invasion of malignant cells, and resistance to chemotherapies. Due to resistance to common therapies, glioma affected patients' survival has not been remarkably improved. ZEB2 (SIP1) is a critical transcriptional regulator with various functions during embryonic development and wound healing that has abnormal expression in different malignancies, including brain tumors. ZEB2 overexpression in brain tumors is attributed to an unfavorable state of the malignancy. Therefore, we aimed to investigate some functions of ZEB2 in two different glioblastoma U87 and U373 cell lines. METHODS: In this study, we investigated the effect of ZEB2 knocking down on the apoptosis, cell cycle, cytotoxicity, scratch test of the two malignant brain tumor cell lines U87 and U373. Besides, we investigated possible proteins and microRNA, SMAD2, SMAD5, and miR-214, which interact with ZEB2 via in situ analysis. Then we evaluated candidate gene expression after ZEB2-specific knocking down. RESULTS: We found that ZEB2 suppression induced apoptosis in U87 and U373 cell lines. Besides, it had cytotoxic effects on both cell lines and reduced cell migration. Cell cycle analysis showed cell cycle arrest in G0/G1 and apoptosis induction in U87 and U373 cell lines receptively. Also, we have found that SAMAD2/5 expression was reduced after ZEB2-siRNA transfection and miR-214 upregulated after transfection. CONCLUSIONS: In line with previous investigations, our results indicated a critical oncogenic role for ZEB2 overexpression in brain glioma tumors. These properties make ZEB2 an essential molecule for further studies in the treatment of glioma cancer.

Simultaneous microRNA-612 restoration and 5-FU treatment inhibit the growth and migration of human PANC-1 pancreatic cancer cells.

Despite the recent advances in the treatment of other cancers, the 5-year survival rate of pancreatic cancer remains under 9 %. Chemotherapy and surgical resection are the most common therapy methods. The regulatory role of microRNAs in different types of cancer has given them therapeutic importance. miR-612 has been downregulated in colorectal, bladder, liver, and some other types of cancer and could be considered a tumor-suppressor miRNA. 5-FU is one of the most common chemotherapeutic agents used in pancreatic cancer treatment, which is used in multiple drug regimens and combinatorial therapy methods. The aim of this study is the evaluation of miR-612 restoration in the PANC-1 cell line and using the tumor-suppressive effect of it in combination with 5-FU on cell growth and migration. MiR-612 mimic was transfected to PANC-1 cells through electroporation. Following the transfection, expression levels of miR-612 and BAX, BCL-2, Caspase-3, MMP9, and PD-L1 genes were measured by qRT-PCR. MTT assay was used to determine the cytotoxicity of miR-612 and 5-FU on PANC-1 cell viability. To confirm MTT results and to evaluate the quantitative effect of apoptosis induction flow cytometry test was used and in order to confirm apoptosis test results and cell cycle arrest evaluation DAPI staining and cell, cycle tests were conducted, respectively. Finally, to assess the inhibitory effect of miR-612 in combination with 5-FU on migration and growth wound healing and colony formation assays were used, respectively. Results demonstrated that miR-612 alongside 5-FU has an important role in the inhibition of migration and growth and also apoptosis induction in PANC-1 cells and could be considered as a supporting agent of chemotherapy and a novel therapeutic modality in pancreatic cancer treatment.

ZEB2 Knock-down Induces Apoptosis in Human Myeloid Leukemia HL-60 Cells.

INTRODUCTION: Acute myeloid leukemia (AML) is the most prevalent type of cancer in the adult hematopoietic system. Conventional therapies are associated with unfavorable side effects in individuals diagnosed with AML. These after-effects with partial remission reflect the urgent need for novel therapeutic approaches for inducing apoptosis, specifically in malignant cells, without affecting other cells. As a transcription factor (TF), ZEB2 (Zinc Finger E-Box Binding Homeobox 2) regulates the expression of specific genes in normal conditions. However, increased expression of ZEB2 is reported in various cancers, especially in AML, which is related to a higher degree of apoptosis inhibition of malignant cells. In this work, the role of ZEB2 in apoptosis inhibition is surveyed through ZEB2 specific knocking-down in human myeloid leukemia HL-60 cells. MATERIALS AND METHODS: Transfection of HL-60 cells was conducted using ZEB2-siRNA at concentrations of 20, 40, 60, and 80 pmol within 24, 48, and 72 h. After determining the optimum dose and time, flow cytometry was used to measure the apoptosis rate. The MTT assay was also utilized to evaluate the cytotoxic impact of transfection on the cells. The expression of candidate genes was measured before and after transfection using qRT-PCR. RESULTS: According to obtained results, suppression of ZEB2 expression through siRNA was associated with the induction of apoptosis, increased pro-apoptotic, and decreased anti-apoptotic gene expression. Transfection of ZEB2-siRNA was also associated with reduced cell proliferation and viability. CONCLUSION: Our study results suggest that ZEB2 suppression in myeloid leukemia cells through apoptosis induction could be a proper therapeutic method.

Mixed methods pilot study of sharing behaviors among waterpipe smokers of rural Lao PDR: implications for infectious disease transmission.

To date, the sharing behaviors associated with the homemade tobacco waterpipe used in rural areas of the Western Pacific Region have not been studied. Evidence from studies of manufactured waterpipes raises the possibility of infectious disease transmission due to waterpipe sharing. The objective of our pilot study in rural Lao People's Democratic Republic (PDR) was to identify and measure the prevalence of waterpipe sharing behaviors. We first conducted ethnographic studies to investigate waterpipe-smoking behaviors. These findings were then used to develop an interviewer-administered household survey that was used in a sampling of waterpipe smokers from three villages of the Luang Namtha province of Lao PDR (n = 43). Sampled waterpipe smokers were predominantly male (90.7%), older (mean age 49, SD 13.79), married (95.4%), farmers (78.6%), and had completed no primary education. Pipes were primarily made from bamboo (92.9%). Almost all (97.6%) smokers were willing to share their pipe with others. At the last time they smoked, smokers shared a pipe with at least one other person (1.2 ± 0.5 persons). During the past week, they had shared a pipe with five other persons (5.2 ± 3.8 persons). The high prevalence of sharing behaviors among waterpipe smokers in rural Southeast Asia raises the possibility that this behavior provides important and unmeasured social network pathways for the transmission of infectious agents.

The role of pseudoephedrine on daytime somnolence in patients suffering from perennial allergic rhinitis (PAR).

BACKGROUND: Allergic rhinitis is one of several inflammatory diseases affecting the nasal mucosa. Cellular inflammation of nasal mucosa is a hallmark of this disease and is characterized by the accumulation of eosinophils and the release of various chemical messengers such as chemokines, cytokines, and histamine. This inflammation of the nose leads to nasal congestion and a reduction in sleep quality, resulting in daytime somnolence. OBJECTIVE: Drugs that significantly reduce the symptoms of nasal congestion also may help in alleviating sleep-related symptoms of allergic rhinitis. Pseudoephedrine is a sympathomimetic amine that is indicated for treatment of nasal congestion associated with allergic rhinitis. Despite relieving nasal congestion, we speculated that, because of pseudoephedrine's well-known stimulant profile, sleep would not be improved. METHOD: Fourteen subjects who met the inclusion criteria were enrolled into a double-blind, placebo-controlled, randomized study to either pseudoephedrine or placebo once per day in the morning, using the traditional crossover design. Skin testing test was performed to ensure a positive response to a relevant perennial allergen and a negative response to a seasonal allergen. Several questionnaires were used to evaluate the patients' sleep-related symptoms, allergic rhinitis symptoms, and quality of life. RESULTS: Our results showed that pseudoephedrine did not have a positive or negative effect on quality of sleep, daytime sleepiness, or daytime fatigue as compared with placebo. Pseudoephedrine did show a statistical significance in improving stuffy nose (P = .0172). With respect to quality of life, pseudoephedrine led to a statistically significant decrease in intimate relationships and sexual activity as compared with the placebo group (P = .0310). CONCLUSION: Our research suggests that sleep quality is not significantly affected by pseudoephedrine. As expected, congestion is reduced, but side effects such as a decline of intimate relationships and sexual activity may interfere with quality of life.

Congestion and sleep impairment in allergic rhinitis.

Allergic rhinitis is a prevalent disease in developed nations, and its prevalence has been increasing throughout the world. Nasal congestion is the most common and bothersome symptoms of rhinitis. Congestion is associated with sleep-disordered breathing and is thought to be a key cause of sleep impairment in individuals with rhinitis. The end result is a decrease in quality of life and productivity and an increase in daytime sleepiness. Treatment with intranasal corticosteroids has been shown to reduce nasal congestion. Data on sleep-related end points from clinical trials of intranasal corticosteroids indicate that this reduction is associated with improved sleep, reduced daytime fatigue, and improved quality of life. Other therapies, such as montelukast, also have a positive influence on congestion and sleep. This review examines nasal congestion and the associated sleep impairment of allergic rhinitis patients. It explores the adverse effects of disturbed sleep on quality of life and how these conditions can be reduced by therapies that decrease congestion.