Mechanism of miRNA-26a on the Proliferation of Pancreatic Tumor Cells by Regulating the Expression of Cycline2

2020 ◽  
Author(s):  
Xiaochun HOU ◽  
Jian XU ◽  
Pei HU
Keyword(s):  
Epithelial Cells ◽  
Cell Lines ◽  
Pancreatic Tumor ◽  
Human Pancreatic Cancer ◽  
Control Group ◽  
Rt Pcr ◽  
Virus Particles ◽  
Apoptotic Protein ◽  
Proliferation Ability ◽  
Apoptosis Proteins

Background: qRT-PCR was used to measure the expression of miRNA-26a in pancreatic epithelial cells (HPDE) and human pancreatic cancer cell lines PANC-1 and MIA PaCa-2. Methods: PANC-1 and MIA PaCa-2 cell lines were infected with lentiviruses to construct PANC-miR-26a and MIA-miR-26a, and RT-PCR was used to detect the infection efficiency. The cell proliferation ability of PANC-miR-26a and MIA-miR-26a were examined by CCK-8 assay, and apoptosis was detected by flow cytometry. Western blotting was used to detect the expressions of CyclinE2 protein and mitochondria-associated apoptotic proteins. Results: miR-26a was expressed in human normal pancreatic epithelial cells (HPDE), and not detected in PANC-1 and MIA PaCa-2; miR-26a was highly expressed in the cell lines PANC-miR-26a and MIA-miR-26a infected by the virus particles. The absorbance values of PANC-miR-26a and MIA-miR-26a were lower than those of NC1 and PANC-1 in control group. The apoptosis rates of PANC-miR-26a and MIA-miR-26a were substantially higher than those of the control group. The overexpression of miR-26a inhibited the expression of the target protein CyclinE2 in PANC-miR-26a and MIA-miR-26a. The expression of the anti-apoptotic protein Bcl-2 was decreased in PANC-miR-26a and MIA-miR-26a, while the expression of the pro-apoptotic protein Bax was increased. Compared with HPDE, miR-26a was down-regulated in PANC-1 and MIA PaCa-2. After overexpression of miR-26a, the proliferation of PANC-1 and MIA PaCa-2 cell lines was weakened. Conclusion: Molecular mechanism is the negative regulation of CyclinE2 by miR-26a as well as the expressions of downstream mitochondrial apoptosis proteins Bcl-2 and Bax.

scholarly journals Application of Magnetic Resonance Image Intelligent Data Acquisition in Ovarian Cancer (Preprint)

2020 ◽  
Author(s):  
Tingting Liang ◽  
Yanlei Dong ◽  
Xinrui Zhao ◽  
Lu Wang ◽  
Hui Xu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Magnetic Resonance ◽  
Epithelial Cells ◽  
Data Acquisition ◽  
Cell Lines ◽  
P53 Protein ◽  
Control Group ◽  
Ovarian Epithelial Cells ◽  
Intelligent Data Acquisition ◽  
Experimental Group

UNSTRUCTURED As a diagnostic method with no radiation, high resolution of soft tissue, and different imaging methods, Magnetic Resonance Imaging (MRI) intelligent data acquisition is more and more widely used in the examination of abdominal, pelvic, and other organ lesions. In order to study the diagnostic effect of multi-mode magnetic resonance intelligent data acquisition on ovarian cancer and the ovarian cancer model modified based on p53-/-+Myc+ASAP1 gene, NSG mice were selected as experimental subjects in this study. 293FT cell lines packaging p53, Myc, and ASAP1(ArfGAP with SH3 domain, Ankyrin repeat and PH domain 1) recombinant lentivirus were inoculated into mouse ovarian epithelial cells to construct mouse ovarian epithelial cell tumor cell lines and their performance was analyzed. According to the different injection cell lines, they were divided into the experimental group and the control group. Tumor samples were collected and the mice were analyzed using immunofluorescence staining and MRI. The results showed that, in the detection of protein expression in genetically modified cell lines, for p53-/-+Myc+ASAP1 fully modified cell lines, the high expression of ASAP1 and Myc functional proteins was detected after the lentivirus containing p53-/-, ASAP1, and Myc were introduced into mouse ovarian epithelial cells, while the expression of p53 protein decreased significantly; after inoculation into mice, it was found that the expression of ASAP1 protein and Myc protein in the experimental group was significantly higher than that in the control group, while the expression of p53 protein in the experimental group was significantly lower than that in the control group, with significant statistical difference; further MRI diagnosis of two groups of mice showed that the ADC (Apparent dispersion coefficient) value of the experimental group was significantly higher than the control group, there were statistically significant differences. Therefore, it was found that p53 gene expression was down-regulated and Myc and ASAPl genes were overexpressed in the tumor tissues and tumor cells formed, and tumor formation differences between the two groups of mice could be obviously found after MRI intelligent data acquisition, which provided experimental basis for early diagnosis of breast cancer in the later clinical stage.

scholarly journals miR-182-5p Promotes Growth in Oral Squamous Cell Carcinoma by Inhibiting CAMK2N1

2018 ◽  
Vol 49 (4) ◽  
pp. 1329-1341 ◽  
Author(s):  
Nan Li ◽  
Chuan-Chuan Nan ◽  
Xue-Yun Zhong ◽  
Jun-Quan Weng ◽  
Hai-Dong Fan ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Oral Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Lines ◽  
Squamous Cell ◽  
Colony Formation ◽  
Cell Counting ◽  
Control Group ◽  
Rt Pcr ◽  
Ki 67

Background/Aims: Emerging evidence suggests that the propagation of oral squamous cell carcinoma (OSCC) is influenced by the abnormal expression of microRNAs (miRNAs). This study aimed to characterize the involvement of miR-182-5p in OSCC by targeting the calcium/ calmodulin-dependent protein kinase II inhibitor CAMK2N1. Methods: miR-182-5p expression was quantified in OSCC tissues and cell lines with reverse transcription polymerase chain reaction (RT-PCR). Cell colony formation, Cell Counting Kit-8 (CCK-8), Ki-67, and nude mouse xenograft assays were used to characterize the role of miR-182-5p in the proliferation of OSCC. A miR-182-5p target gene was identified with western blotting, RT-PCR, and luciferase activity assays. OSCC patient survival based on CAMK2N1 expression was also analyzed. Results: miR-182-5p was up-regulated in in vitro cell lines and in vivo clinical OSCC samples. CCK-8, colony formation, and Ki-67 assays revealed that miR-182-5p promoted the growth and proliferation of OSCC cells. miR-182-5p directly targeted CAMK2N1, as evidenced by luciferase assays and target prediction algorithms. CAMK2N1 operated as a tumor suppressor gene in patients with OSCC. Down-regulating miR-182-5p expression in the CAL-27 cell line restored CAMK2N1-mediated OSCC cell proliferation. miR-182-5p expression inhibited the activation of AKT, ERK1/2, and NF-κB. Mice injected with CAL-27 cells transfected with miR-182-5p-inhibitor demonstrated a significant increase in tumor size and weight and increased CAMK2N1 mRNA and protein expression compared with the miR-negative control group. Conclusion: The miR-182-5p-CAMK2N1 pathway can be potentially targeted to regulate the proliferation of OSCC cells.

scholarly journals Oral Glutamine Supplement Inhibits Ascites Formation in Peritoneal Carcinomatosis Mouse Model

2013 ◽  
Vol 2013 ◽  
pp. 1-4
Author(s):  
Ming-Jen Chen ◽  
Tsang-En Wang ◽  
Shu-Jung Tsai ◽  
Ching-Chung Lin ◽  
Chia-Yuan Liu ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Serum Albumin ◽  
Peritoneal Carcinomatosis ◽  
Ascitic Fluid ◽  
Pancreatic Tumor ◽  
Preclinical Study ◽  
Human Pancreatic Cancer ◽  
Control Group ◽  
Cell Mediated Immunity ◽  
Ascites Formation

Background. Peritoneal carcinomatosis (PC) accompanied with ascites formation causes several distressing symptoms, resulting in poor quality of life.Methods. Twenty BALB/c nude mice generated by direct orthotopic injection of human pancreatic cancer PANC-1 cells were randomized to receive either a stock laboratory diet or a stock diet supplemented with glutamine. Half of the mice were sacrificed at day 76 to measure the amount of ascitic fluid and pancreatic tumor volume. The remaining mice were subject to survival analysis. Serum albumin levels were estimated every 2 weeks.Results. At day 76, the average amount of ascitic fluid measured in the control group was1.2±0.3 mL compared to0.5±0.5 mL from the glutamine-supplemented mice (P=0.045). The volume of pancreatic tumor was2.60±0.8 cm3in the control group and1.98±1.3 cm3in glutamine-supplemented mice (P=0.39). The mean survival time of glutamine-supplemented mice was prolonged from87±4to101±2days (P=0.0024). Mean serum albumin levels were higher in the glutamine-supplemented group.Conclusions. This preclinical study showed that oral supplementation of glutamine may provide ascites-reducing activity in pancreatic cancer patients with PC, via a cell-mediated immunity-independent mechanism.

scholarly journals Role of MMP-9 in Epithelial-Mesenchymal Transition of Thyroid Cancer

2020 ◽  
Author(s):  
Yuanchun Li ◽  
Jing He ◽  
Feng Wang ◽  
Xin Wang ◽  
Fan Yang ◽  
...  
Keyword(s):  
Cell Lines ◽  
Migration And Invasion ◽  
Control Group ◽  
Thyroid Cell ◽  
Cell Marker ◽  
Rt Pcr ◽  
Invasion And Metastasis ◽  
Thyroid Cell Line ◽  
Tgf Β1 ◽  
E Cadherin

Abstract Background The purpose of this study is to explore the role and mechanism of MMP-9 in the EMT process of thyroid cancer (TC), so as to provide a basis for clinical exploration of invasion and metastasis process of TC, looking for biological markers of tumor metastasis and molecular intervention therapy.Methods Western Blot and RT-PCR were employed to detect the expression of MMP-9 in human normal thyroid cell line HT-ori3 and human TC cell lines IHH-4 (PTC), FTC-133 and 8505C. Expression levels of EMT-related markers: epithelial cell marker E-cadherin and stromal cell marker Vimentin in TGF-1-induced TC cell lines were detected by Western Blot and RT-PCR, respectively. The effects of MMP-9 down-regulation on cell invasion and metastasis were investigated by wound-healing assay and cell invasion experiment.Results The protein and mRNA expression levels of MMP-9 in TC cell lines were increased compared with human normal thyroid cell line HT-ori3. When TGF-β1 was added, the expression of EMT and Vimentin increased while the expression of E-cadherin decreased. Compared with the control group, the TC cells stably transfected with MMP-9 shRNA showed inhibited EMT, decreased Vimentin expression and increased E-cadherin expression. The induction of TGF-β1 did not promote the occurrence of EMT in TC cells which were stably transformed with MMP-9 shRNA. The addition of TGF-β1 to TC cells increased the ability of the cells to migrate and invade. Compared with the control group, the migration and invasion ability of TC cells stably transfected with MMP-9 shRNA was significantly reduced, and the induction of TGF-β1 could not restore the migration and invasion ability of cells without MMP-9.Conclusions In conclusion, we found that MMP-9 can be used as a biomarker for TC, which can promote the EMT process of TGF-β1 induced TC, and thus affecting the cell migration and invasion ability.

(EB) Virus: Latency and Expression in Transplanted and Cultured Human Cells

1971 ◽  
Vol 29 ◽  
pp. 368-369
Author(s):  
B. G. Uzman ◽  
M. M. Kasac ◽  
H. Saito ◽  
A. Krishan
Keyword(s):  
Cell Lines ◽  
Primary Cultures ◽  
Virus Particles ◽  
Barr Virus ◽  
Virus Latency ◽  
Virus Surveillance ◽  
Cultured Human Cells ◽  
Epstein Barr ◽  
Serial Transplantation ◽  
Tubular Arrays

In conjunction with the cultivation and transplantation of cells from human tumors by the Programs of Microbiology and Immunogenetics, virus surveillance by electron microscopy has been routinely employed. Of particular interest in this regard have been 3 cell lines cultured from lymph nodes or spleen of 2 patients with Hodgkin's disease and 1 patient with Letterer-Siwe's disease. Each of these cell lines when transplanted in Syrian hamster neonates conditioned with anti-lymphocyte serum grew as serially transplantable tumors; from such transplants of the 3 cell lines cell cultures were retrieved.Herpes type virus particles (Figs. 1, 2, 3) were found in the primary cultures of all three lines, in frozen thawed aliquots of same, and in cultures retrieved from their tumors growing by serial transplantation in hamsters. No virus was detected in sections of 25 of the serially transplanted tumors. However, in 10 such tumors there were repeated instances of tubular arrays in the cisternae of the endoplasmic reticulum (Fig. 4). On serologic examination the herpes virus was shown to be the Epstein-Barr virus.

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Immunohistochemical Analysis ◽  
Specific Protein ◽  
Platelet Membrane ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Megakaryocytic Cell

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

scholarly journals Comparative evaluation of osteogenic differentiation potential of stem cells derived from dental pulp and exfoliated deciduous teeth cultured over granular hydroxyapatite based scaffold

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Manal Nabil Hagar ◽  
Farinawati Yazid ◽  
Nur Atmaliya Luchman ◽  
Shahrul Hisham Zainal Ariffin ◽  
Rohaya Megat Abdul Wahab
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
3D Culture ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Osteogenic Potential ◽  
Control Group ◽  
Rt Pcr ◽  
Differentiation Potential

Abstract Background Mesenchymal stem cells isolated from the dental pulp of primary and permanent teeth can be differentiated into different cell types including osteoblasts. This study was conducted to compare the morphology and osteogenic potential of stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC) in granular hydroxyapatite scaffold (gHA). Preosteoblast cells (MC3T3-E1) were used as a control group. Methodology The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase assay was used to compare the osteoblastic differentiation of these cells (2D culture). Then, cells were seeded on the scaffold and incubated for 21 days. Morphology assessment using field emission scanning electron microscopy (FESEM) was done while osteogenic differentiation was detected using ALP assay (3D culture). Results The morphology of cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. In RT-PCR study, DPSC and SHED expressed GAPDH, CD73, CD105, and CD146 while negatively expressed CD11b, CD34 and CD45. FESEM results showed that by day 21, dental stem cells have a round like morphology which is the morphology of osteoblast as compared to day 7. The osteogenic potential using ALP assay was significantly increased (p < 0.01) in SHED as compared to DPSC and MC3T3-E1 in 2D and 3D cultures. Conclusion gHA scaffold is an optimal scaffold as it induced osteogenesis in vitro. Besides, SHED had the highest osteogenic potential making them a preferred candidate for tissue engineering in comparison with DPSC.

scholarly journals Down-regulation LncRNA-SNHG15 contributes to proliferation and invasion of bladder cancer cells

BMC Urology ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Aldhabi Mokhtar ◽  
Chuize Kong ◽  
Zhe Zhang ◽  
Yan Du
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Tumor Stage ◽  
Rt Pcr ◽  
Down Regulation ◽  
Bladder Cancer Cells ◽  
Cancer Tissues ◽  
Proliferation And Invasion

Abstract Objectives The aim of this study was to investigate the effect of lncRNA-SNHG15 in bladder carcinoma using cell lines experiments and the relationship between clinical characteristics and lncRNA-SNHG15 expression was analyzed. Methods Bladder cancer tissues and near-cancer tissues were collected. The real-time PCR (RT-PCR) was used to detect the expression of lncRNA-SNHG15 in tissues and cell lines. The expression of lncRNA-SNHG15 was downregulated by interference (siRNA), as detected by RT-PCR, that was used to determine the efficiency of the interference. CCK-8 and Transwell assays were used to evaluate the effect of lncRNA-SNHG15 on the proliferation and invasion capability of bladder cancer cells. The t-test was used for Statistical analyses, which were carried out using the Statistical Graph pad 8.0.1.224 software. Result The expression of lncRNA-SNHG15 was up regulated in 5637, UMUC3 and T24 cell lines compared with corresponding normal controls (P < 0.05). Up regulation was positively related to tumor stage (P = 0.015). And tumor size (P = 0.0465). The down-regulation of lncRNA-SNHG15 with siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion. Conclusion This study showed that lncRNA-SNHG15 is overexpressed in bladder cancer tissues and (5637, UMUC3 T24) cell lines. Up regulation was positively related to tumor stage (P = 0.015), and tumor size (P = 0.0465). Down-regulation of lncRNA-SNHG15 by siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion, indicating a potential molecular target for future tumor targeted therapy.

scholarly journals Anticancer Potential of Biogenic Silver Nanoparticles: A Mechanistic Study

Pharmaceutics ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 707
Author(s):  
Mohd Shahnawaz Khan ◽  
Alya Alomari ◽  
Shams Tabrez ◽  
Iftekhar Hassan ◽  
Rizwan Wahab ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Cancer Cells ◽  
Cell Lines ◽  
Human Life ◽  
Mechanistic Study ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Cell Viability Assay ◽  
Hct 116 ◽  
Mcf 7

The continuous loss of human life due to the paucity of effective drugs against different forms of cancer demands a better/noble therapeutic approach. One possible way could be the use of nanostructures-based treatment methods. In the current piece of work, we have synthesized silver nanoparticles (AgNPs) using plant (Heliotropiumbacciferum) extract using AgNO3 as starting materials. The size, shape, and structure of synthesized AgNPs were confirmed by various spectroscopy and microscopic techniques. The average size of biosynthesized AgNPs was found to be in the range of 15 nm. The anticancer potential of these AgNPs was evaluated by a battery of tests such as MTT, scratch, and comet assays in breast (MCF-7) and colorectal (HCT-116) cancer models. The toxicity of AgNPs towards cancer cells was confirmed by the expression pattern of apoptotic (p53, Bax, caspase-3) and antiapoptotic (BCl-2) genes by RT-PCR. The cell viability assay showed an IC50 value of 5.44 and 9.54 µg/mL for AgNPs in MCF-7 and HCT-116 cell lines respectively. We also observed cell migration inhibiting potential of AgNPs in a concentration-dependent manner in MCF-7 cell lines. A tremendous rise (150–250%) in the production of ROS was observed as a result of AgNPs treatment compared with control. Moreover, the RT-PCR results indicated the difference in expression levels of pro/antiapoptotic proteins in both cancer cells. All these results indicate that cell death observed by us is mediated by ROS production, which might have altered the cellular redox status. Collectively, we report the antimetastasis potential of biogenic synthesized AgNPs against breast and colorectal cancers. The biogenic synthesis of AgNPs seems to be a promising anticancer therapy with greater efficacy against the studied cell lines.

scholarly journals Chimeric Measles Virus (MV/RSV), Having Ectodomains of Respiratory Syncytial Virus (RSV) F and G Proteins Instead of Measles Envelope Proteins, Induced Protective Antibodies against RSV

Vaccines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 156
Author(s):  
Akihito Sawada ◽  
Takashi Ito ◽  
Yoshiaki Yamaji ◽  
Tetsuo Nakayama
Keyword(s):  
Respiratory Syncytial Virus ◽  
G Proteins ◽  
Cell Lines ◽  
Measles Virus ◽  
Pathological Finding ◽  
Control Group ◽  
Protein Coding ◽  
Virus Growth ◽  
Protective Antibodies ◽  
Syncytial Virus

In our previous study, fusion (F) or glyco (G) protein coding sequence of respiratory syncytial virus (RSV) was inserted at the P/M junction of the measles AIK-C vector (MVAIK), and the recombinant measles virus induced protective immune responses. In the present study, the ectodomains of measles fusion (F) and hemagglutinin (HA) proteins were replaced with those of RSV F and G proteins, and a chimeric MV/RSV vaccine was developed. It expressed F and G proteins of RSV and induced cytopathic effect (CPE) in epithelial cell lines (Vero, A549, and HEp-2 cells), but not in lymphoid cell lines (B95a, Jurkat, and U937 cells). A chimeric MV/RSV grew similarly to AIK-C with no virus growth at 39 °C. It induced NT antibodies against RSV in cotton rats three weeks after immunization through intramuscular route and enhanced response was observed after the second dose at eight weeks. After the RSV challenge with 106 PFU, significantly lower virus (101.4±0.1 PFU of RSV) was recovered from lung tissue in the chimeric MV/RSV vaccine group than in the MVAIK control group with 104.6±0.2 PFU (p < 0.001) and no obvious inflammatory pathological finding was noted. The strategy of ectodomain replacement in the measles virus vector is expected to lead to the development of safe and effective vaccines for other enveloped viruses.

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