The main drug studied in this article is gemcitabine, which is mainly used in the treatment of patients with advanced pancreatic cancer. However, if patients develop drug resistance to such drugs, it will inevitably affect the quality of life and even life of patients. At present, the research on the mechanism of gemcitabine resistance to pancreatic cancer is still unclear. The authors first constructed a gemcitabine-resistant cell line and compared the expression profile of circular RNA between drug-resistant and normal cell lines by whole transcriptome sequencing . Through the bio-information analysis, 10 circular RNAs of interest and differential expression were quickly found, and the verification results were consistent with the sequencing results. Two circular RNAs with high expression in gemcitabine-resistant patients were selected and verified in 40 clinical plasma samples. The expression patterns of the two were unchanged. The circular RNA and miRNA network maps show the miRNAs that both may bind. Silencing and overexpression experiments confirmed that these circular RNAs may affect the function of gemcitabine resistant cell lines. Collectively, this article provides a new molecular marker for studying gemcitabine resistance.
Journal: frontiers in Pharmacology
Impact factor: 3.8
Service content: Whole transcriptome sequencing (provided by Cloud Sequence Bio)
Samples: 1. Gemcitabine-resistant pancreatic cancer cell line and untreated pancreatic cancer cell line, sample size: 3 vs 3
2. Drug-resistant patient plasma and healthy human plasma, sample size: 20 vs 20
Published time: June 2018
Article content:
First, the authors tested the activity of cells treated with different concentrations of gemcitabine by MTT assay. After determining the semi-inhibitory concentration, the proliferation curve was used to confirm that the drug can inhibit cell proliferation. In addition, the authors also examined the expression of the MDR1 gene, a marker that is highly expressed in most resistant cell lines, and found that it is highly expressed in the resistant PANC-1 cell line. The above results confirmed that the drug-resistant cell line was successfully constructed.
6. Cyclic RNA inhibits the sensitivity of drug-resistant pancreatic cancer cells to drugs and promotes drug resistance in pancreatic cancer cells of the control group.
To study the previously selected loops, the authors constructed siRNAs and performed interference experiments in drug-resistant cell lines. It was found that the loops can promote the sensitivity of drug-resistant cell lines to drugs and affect apoptosis. Overexpression experiments confirmed that the two loops affected cell resistance in the control cancer cells.
to sum up:
In general, this article first validates the function of gemcitabine in pancreatic cancer cells, and then constructs a full-transcriptome sequencing of drug-resistant and normal pancreatic cancer cell lines, and screens 10 of the most differentially expressed rings from the sequencing results. RNA. Two low-throughput validations of clinical samples yielded two circular RNAs that are highly expressed in drug-resistant cell lines, predicting their binding to miRNAs. Finally, the two loops were knocked down and overexpressed, respectively, confirming that both are involved in the process of cell resistance.
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