Biology Case Study - 983 Words

Materials and methods The levels of microRNA-22-3p (miR-22-3p) and TP63 were analyzed by RT-qPCR and/or Western blot. Cell proliferation was measured using MTT assay. Cell invasion and migration were evaluated using Transwell assay chambers. Prediction of a regulatory relationship between microRNA-22-3p and 3-UTR of TP63 mRNA was performed by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. OC cell lines Lung cancer cell lines H292, PC-9, CL1-5, A549 and normal NHBE cells were used in this study. All of Lung cancer and normaL ceLL Lines - were provided by ATCC. H292, PC-9, CL1-5, A549 and NHBE cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS). MiRNA and RNA interference†¦show more content†¦The experiments were repeated thrice. Cell migration and invasion assays In vitro cell migration and invasion were examined using the Boyden Chamber Assays. For the cell migration assay, 30000 cells in 100 ÃŽ ¼l DMEM medium devoid of fetal bovine serum (FBS) and a given concentration of cetuximab were cultured on a polycarbonate membrane insert coated with fibronectin in a Boyden Chamber (Essen BioScience, UK). In the lower chamber, 500 ÃŽ ¼l DMEM with 10% FBS was added as chemoattractant. After the cells were incubated for 6 h at 37  °C in a 5% CO2 atmosphere, the insert was rinsed using PBS and cells on the upper side of the insert were gently removed using a cotton swab. Subsequently, cells that adhered to the lower side were fixed with methanol, stained with crystal violet solution and counted under a microscope in five predetermined fields (Ãâ€"200). All assays were independently repeated at least thrice. The same protocol as described above was used for the cell invasion assay, except that the transwell filter membranes were pre-coated with 24 à Ž ¼g/ÃŽ ¼l matrigel (RD Systems, Inc., Minneapolis, MN, USA) and the cells were incubated for 8 h at 37  °C in a 5% CO2 atmosphere. RNA isolation, reverse transcription and RT–qPCR RNA was extracted from the OC cells using the Trizol reagent. For miR-22-3p, RNA was transcribed into complementary DNA and amplified using miRNA PrimeScript RT Enzyme Mix kit according to the manufacturer’s instructionsShow MoreRelatedBiology Case Study1218 Words   |  5 Pagesconvenient for our sample study since heart attack disease has been found to start developing in as early a range as 20-39 years (Mozaffarian 2015). Some interesting things to note about our population sample is the changes overtime they had from 1950 to 1962. The weight change decreased about 1.4 pounds as did the serum cholesterol by 4.5 mg% and BMI by 0.2298964. 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