Identification of best housekeeping genes for the normalization of RT-qPCR in human cell lines

Acta Histochem. 2021 Nov 30;124(1):151821. doi: 10.1016/j.acthis.2021.151821. Online ahead of print.ABSTRACTThe identification of the best reference gene is a critical step to evaluate the relative change in mRNA expression of a target gene by RT-qPCR. In this work, we evaluated nineteen genes of different functional classes using Real Time Human Reference Gene Panel (Roche Applied Sciences), to identify the internal housekeeping genes (HKGs) most suitable for gene expression normalization data in human cell lines. Normal cell lines CCD-19LU (lung fibroblast), HEK-293 (epithelial cell of embryonic kidney), WI-26 VA4 (lung fibroblast), and human cancer cells, BT-549 (breast cancer), Hs 578T (breast cancer), MACL-1 (breast cancer), HeLa (cervical carcinoma), U-87 MG (glioblastoma/astrocytoma), RKO-AS45-1 (colorectal carcinoma), and TOV-21G (ovarian adenocarcinoma) were cultivated according to manufacturer's protocol. Twelve candidate reference genes were commonly expressed in five cell lines (CCD-19Lu, HEK-293, RKO-AS45-1, TOV-21G, and U-87 MG). To verify the expression stability, we used the RefFinder web tool, which integrates data from the computational programs Normfinder, BestKeeper, geNorm, and the comparative Delta-Ct method. The ACTB was the most stable reference gene to the CCD-19Lu and HEK-293 cells. The best combination of HKGs for the RKO-AS45-1 and TOV-21G cell lines were B2M/GAPDH and PBGD/B2M, respectively. For the U-87 MG cells, GAPDH and IPO8 were the most suit...
Source: Acta Histochemica - Category: Biochemistry Authors: Source Type: research

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ConclusionThe DCE-MRI radiomics model, especially image features extracted from the delayed phases, can help improve the performance in predicting TILs. The radiomics nomogram is effective in predicting TILs in breast cancer.Key Points•Radiomics features extracted from DCE-MRI, especially delayed phase images, help predict TIL levels in breast cancer.•We developed a nomogram based on MRI to predict TILs in breast cancer that achieved the highest AUC of 0.950.
Source: European Radiology - Category: Radiology Source Type: research
ConclusionWe observed strong reproducibility in the assessment of PI-QUAL between two radiologists with high expertise in prostate mpMRI. At present, PI-QUAL offers clinicians the only available tool for evaluating and reporting the quality of prostate mpMRI in a systematic manner but further refinements of this scoring system are warranted.Key Points•Inter-reader agreement for each single Prostate Imaging Quality (PI-QUAL) score (i.e., PI-QUAL 1 to PI-QUAL 5) was strong, with weighted kappa = 0.85 (95% confidence intervals: 0.51 – 1) and percent agreement = 84%.• Interobserver  agreement was strong wh...
Source: European Radiology - Category: Radiology Source Type: research
This study was designed to compare the detection of subtle lesions (calcification clusters or masses) when using the combination of digital breast tomosynthesis (DBT) and synthetic mammography (SM) with digital mammography (DM) alone or combined with DBT.MethodsA set of 166 cases without cancer was acquired on a DBT mammography system. Realistic subtle calcification clusters and masses in the DM images and DBT planes were digitally inserted into 104 of the acquired cases. Three study arms were created: DM alone, DM with DBT and SM with DBT. Five mammographic readers located the centre of any lesion within the images that s...
Source: European Radiology - Category: Radiology Source Type: research
AbstractObjectivesTo evaluate the correlation between liver enhancement on hepatobiliary phase and liver function parameters in a multicenter, multivendor study.MethodsA total of 359 patients who underwent gadoxetic acid –enhanced MRI using a standardized protocol with various scanners within a prospective multicenter phase II trial (SORAMIC) were evaluated. The correlation between liver enhancement on hepatobiliary phase normalized to the spleen (liver-to-spleen ratio, LSR) and biochemical laboratory parameters, clinical findings related to liver functions, liver function grading systems (Child-Pugh and Albumin-Bili...
Source: European Radiology - Category: Radiology Source Type: research
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Source: Journal of the American Academy of Physician Assistants - Category: Primary Care Tags: Quick Recertification Series Source Type: research
The number of people living with chronic immunosuppression is increasing in the United States. Patients with HIV, those who have had bone marrow or solid organ transplants, and patients taking biologics for autoimmune diseases are at increased risk for skin cancer. Skin cancer in these patients is more aggressive and more likely to metastasize and cause death. Medications and individual risk factors such as sex, age, and ethnicity are independent risk factors for the development of skin cancer. Routine screening and aggressive treatment of actinic keratoses and nonmelanoma skin cancers can reduce patients' skin cancer burd...
Source: Journal of the American Academy of Physician Assistants - Category: Primary Care Tags: CME: Primary Care Source Type: research
Acta Histochem. 2021 Nov 30;124(1):151821. doi: 10.1016/j.acthis.2021.151821. Online ahead of print.ABSTRACTThe identification of the best reference gene is a critical step to evaluate the relative change in mRNA expression of a target gene by RT-qPCR. In this work, we evaluated nineteen genes of different functional classes using Real Time Human Reference Gene Panel (Roche Applied Sciences), to identify the internal housekeeping genes (HKGs) most suitable for gene expression normalization data in human cell lines. Normal cell lines CCD-19LU (lung fibroblast), HEK-293 (epithelial cell of embryonic kidney), WI-26 VA4 (lung ...
Source: Acta Histochemica - Category: Biochemistry Authors: Source Type: research
Li Liu, Jiajing Lin and Hongying He* Department of Obstetrics and Gynecology, Liuzhou Worker’s Hospital, Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, China Background and Objective: Endometrial cancer (EC) is a common gynecological malignancy worldwide. Despite advances in the development of strategies for treating EC, prognosis of the disease remains unsatisfactory, especially for advanced EC. The aim of this study was to identify novel genes that can be used as potential biomarkers for identifying the prognosis of EC and to construct a novel risk stratification using these genes. Me...
Source: Frontiers in Genetics - Category: Genetics & Stem Cells Source Type: research
Markus Hartl* and Rainer Schneider Center of Molecular Biosciences (CMBI), Institute of Biochemistry, University of Innsbruck, Innsbruck, Austria The neuronal proteins GAP43 (neuromodulin), MARCKS, and BASP1 are highly expressed in the growth cones of nerve cells where they are involved in signal transmission and cytoskeleton organization. Although their primary structures are unrelated, these signaling proteins share several structural properties like fatty acid modification, and the presence of cationic effector domains. GAP43, MARCKS, and BASP1 bind to cell membrane phospholipids, a process reversibly regulate...
Source: Frontiers in Oncology - Category: Cancer & Oncology Source Type: research
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