Clone 5-1-111/26/2022 ![]() ![]() Also, because the data are all numerical and standardized, they are easily combined into interactive transcript abundance indices (ITAI). This leads to synergistically increasing value of data over time as data accumulate. With StaRT-PCR, data from all experiments may be directly compared in the same database. In an effort to identify biomarkers that diagnose BC with more accuracy than cytomorphologic criteria, this laboratory developed and has employed a highly quantitative, quality controlled RT-PCR method, Standardized RT (StaRT)-PCR. It is reasonable to expect that improved mechanistic understanding of BC will lead to reduced death rate through more effective prevention and treatment regimens. ![]() Specifically, they provide experimental confirmation that a line with slope of 7,000 on the p21 versus bivariate plot quantifies the difference between normal and malignant BEC at the level of transcript abundance.īronchogenic carcinoma (BC) is the leading cause of cancer-related death in the United States and most industrialized nations. These results, likely due to increased p21-mediated down-regulation of E2F1 TA at the G1/S phase transition, are consistent with our hypothesis. This change was associated with an approximate 2-fold increase in population doubling time to 38.4 hours (from 22.3 hrs), resumption of contact-inhibition, and reduced dividing cell fraction as measured by flow cytometric DNA analysis. ![]() Specifically, each transfectant was capable of only 15 population doublings compared with unlimited population doublings for parental A549. These changes in regulation of cell cycle control genes were associated with restoration of cell proliferation control. For each clone, the increase in p21 transcript abundance (TA) was associated with increased total p21 protein level, more than 5-fold reduction in E2F1 TA, and 10-fold reduction in the /p21 ITAI to a value below the cut-off threshold. Resultsįollowing transfection of p21, four A549/p21 clones with stable two-fold up-regulated p21 expression were isolated and expanded. In order to test the hypothesis, a p21 expression vector was transfected into the A549 human bronchogenic carcinoma cell line, which has low constitutive p21 TA expression relative to normal BEC. The hypothesis was tested that experimental elevation of p21 in a malignant BEC line would decrease the value of the /p21 ITAI to a level below this line, resulting in loss of immortality and limited cell population doubling capacity. This study was an effort to better quantify, at the transcript abundance level, the difference between normal and malignant BEC. This cut-off is represented by a line with a slope of 7,000 on a bivariate plot of p21 versus, with malignant BEC above the line and normal BEC below the line. In previous studies, an interactive transcript abundance index (ITAI) comprising three cell cycle control genes, /p21 accurately distinguished normal from malignant bronchial epithelial cells (BEC), using a cut-off threshold of 7,000. Deregulated cell cycle progression and loss of proliferation control are key properties of malignant cells. ![]()
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