Comprehensive Oncology Drug Research Program
Based on TCGA, GEO big data or Creative Proteomics database or customer samples, new functional genes that have an anti-tumor effect and have not been reported in the studied tumor are screened for new functional genes. Cells, A variety of experimental verifications at the in vivo level, and explore the potential mechanism of the screened genes, to provide a preliminary data basis for the determination of the target of antitumor Chinese medicine drugs and biomarkers.
The whole gene expression profiling chip was used to analyze the changes in the gene expression of Chinese medicine monomers with antitumor effects on tumor cell lines. Among these differential genes, down-regulated genes stimulated by Chinese medicine monomers are likely to promote tumor cell proliferation. These genes can be suppressed by the traditional Chinese medicine monomer, thereby inhibiting cell proliferation.
Combined with the patent database of key genes for CP gene diseases and the high-content functional gene screening platform (HCS), those genes that were down-regulated after treatment of these traditional Chinese medicine monomers were screened out that have not been reported in the tumors studied and played a role in tumor cell proliferation Important regulatory role of downstream target genes. Complete the functional experiments of the target gene at the level of cells and animals, and clarify the molecular mechanism of these key genes through supplementary research programs, and explore the mechanism of action of the traditional Chinese medicine monomer.
1. Chinese medicine monomer anti-tumor function pre-experiment: According to the Chinese medicine monomer provided by the customer, IC50 measurement was performed on tumor cell lines to determine the specific concentration, conditions and duration, and apoptosis cell cycle scratch healing was formed by MTT clone Other experiments verify the effectiveness of traditional Chinese medicine monomers.
2. Gene chip screening for differentially expressed genes after TCM monomer treatment: Cells were treated according to the established conditions of TCM monomer application in the pre-experiment, total RNA was extracted, and the whole gene expression profile chip test was performed with the control group, and the differences were screened through informatics analysis Genes with significant expression were selected, and about 25-40 down-regulated differential genes were selected for qPCR verification, and 20 candidate differential genes were identified.
3. Preliminary screening of functional target genes: 20 candidate gene shRNA lentiviruses were used to infect target cells, and each gene was replicated in 3 wells to reduce the expression of candidate genes endogenously expressed in the target cells. Cellomics High Content Functional Screening (HCS) On the platform, the infected cells were counted for 4 to 5 days, and the cell proliferation of the cells with down-regulated candidate gene expression compared with control cells was observed. About 1-2 target genes that had an effect on cell proliferation were initially selected.
4. Perform a complete cell functional study on one of the functional target genes screened: verify the knockdown efficiency of the target gene B in cells by qPCR and WB, and perform MTT / cell cycle / apoptosis / BrdU on tumor cell lines / Clone formation and other functional experiments in the direction of proliferation.
5. In vivo functional studies of functional target genes: prepare target gene knockdown experimental groups or control tumor cells, inject nude mice, feed nude mice to tumor growth, record animal weight, tumor size curve, and kill The tumors were collected and weighed to process the data results.
6. Research on target gene action mechanism: through cellular immunofluorescence, Patharray downstream pathway analysis, downstream classical pathway WB verification, Co-IP mass spectrometry analysis of interaction proteins, dryness detection, aging detection, DNA methylation detection, acetylation detection Complementary research schemes such as ubiquitination detection and autophagy mechanism verification were performed to verify the possible pathogenic molecular mechanisms of target genes.
7. Experimental analysis of target gene and drug function recovery: construct target gene over-expressing lentivirus, WB to verify the target gene expression efficiency, infect the target gene over-expressing lentivirus in tumor cell lines, and co-process with Chinese medicine monomers, etc. through CCK8 Experiment to detect cell proliferation and observe whether the effect is weakened.
Creative Proteomics leads a rich team that uses the most rigorous experimental solutions and internationally recognized experimental techniques for Personalized research plan for Comprehensive Oncology Drug Research Program to obtain your customized information. Use the most accurate experimental results to help you better carry out tumor-related scientific research.
Mature and reliable Personalized research plan for Comprehensive Oncology Drug Research Program platform: rich project experience, participate in major projects, and publish high-level cooperation results.
The analysis contents and methods based on high-quality data analysis, pre-research and verification are indeed mature and feasible.
Please contact us to find out how we can help you achieve the next research breakthrough.