Mechanism Exploration and Screening for Drug Resistance Induced by SAM-RNA in Breast Cancer
Session Number
Project ID: BIO 11
Advisor(s)
Tong-Chuan He, University of Chicago Biological Science Division
Hui Zhang, University of Chicago Biological Science Division
Discipline
Biology
Start Date
17-4-2024 9:20 AM
End Date
17-4-2024 9:35 AM
Abstract
Some patients who are initially sensitive to such chemotherapy and endocrine therapy quickly develop acquired drug resistance. We aim to explore the molecular changes and potential regulatory mechanisms during the acquisition of chemotherapy and endocrine therapy resistance in breast cancer, or further to find molecular indicators that can be used to early predict the occurrence of drug resistance or efficacy evaluation. Various biological assays, such as crystal violet staining, flow cytometry, Hoechst staining, and real-time PCR will be utilized to comprehensively evaluate the effects of SAM-RNA overexpression on cellular processes and functions associated with breast cancer cell lines. Signal pathways involved in tamoxifen-resistant and paclitaxel-resistant will be analyzed by Exon sequencing and Transcriptome sequencing. In vivo tests will be done by using mouse tumorigenesis models. Molecular cloning was used to construct a retroviral expression library containing 19 random bases (N19), namely short artificial modulatory RNA (SAM-RNA) synthesized by chemical approach and amplified by PCR. The library diversity and properties were evaluated by PCR, whole genome sequencing. Based on N19 library results, stable cell lines expressing SAM RNA1 and SAM RNA7 were constructed by pseudotyped retroviral vectors. Tamoxifen-resistant and paclitaxel-resistant cell lines expressing SAM RNA1 and SAM RNA7 were constructed.
Mechanism Exploration and Screening for Drug Resistance Induced by SAM-RNA in Breast Cancer
Some patients who are initially sensitive to such chemotherapy and endocrine therapy quickly develop acquired drug resistance. We aim to explore the molecular changes and potential regulatory mechanisms during the acquisition of chemotherapy and endocrine therapy resistance in breast cancer, or further to find molecular indicators that can be used to early predict the occurrence of drug resistance or efficacy evaluation. Various biological assays, such as crystal violet staining, flow cytometry, Hoechst staining, and real-time PCR will be utilized to comprehensively evaluate the effects of SAM-RNA overexpression on cellular processes and functions associated with breast cancer cell lines. Signal pathways involved in tamoxifen-resistant and paclitaxel-resistant will be analyzed by Exon sequencing and Transcriptome sequencing. In vivo tests will be done by using mouse tumorigenesis models. Molecular cloning was used to construct a retroviral expression library containing 19 random bases (N19), namely short artificial modulatory RNA (SAM-RNA) synthesized by chemical approach and amplified by PCR. The library diversity and properties were evaluated by PCR, whole genome sequencing. Based on N19 library results, stable cell lines expressing SAM RNA1 and SAM RNA7 were constructed by pseudotyped retroviral vectors. Tamoxifen-resistant and paclitaxel-resistant cell lines expressing SAM RNA1 and SAM RNA7 were constructed.