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Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells
Published on: February 21, 2014
Chemoresistance‑related long non‑coding RNA expression profiles in human breast cancer cells
Lei Huang1, Lihua Zeng2, Jiahui Chu1
1Department of Breast Surgery, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Abstract:
Breast cancer is the most commonly diagnosed cancer and the leading cause of cancer death in females worldwide. Chemoresistance has been a major reason for the drug therapy failure. The present study performed a microarray analysis between MCF‑7 and MCF‑7/adriamycin (ADR) cells, and intended to identify long non‑coding (lnc)RNA expression character in drug resistant breast cancer cells. MCF‑7/ADR cells were induced from MCF‑7 cells via pulse‑selection with doxorubicin for 4 weeks, and the resistance to doxorubicin of ADR cells was confirmed by MTT assay. Microarray analysis was performed between MCF‑7 and MCF‑7/ADR cells. Total RNA was extracted from the two cell lines respectively and was transcribed into cDNA. The results of the microarray were verified by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Gene Ontology (GO) and pathways analysis were conducted to enrich the dysregulated lncRNAs presented in the microarray results. Compared to the MCF‑7 cells, 8,892 lncRNAs were differentially expressed in MCF/ADR cells (absolute fold‑change >2.0). A total of 32 lncRNAs were selected for RT‑qPCR by fold‑change filtering, standard Student's t‑test, and multiple hypothesis testing. Among the dysregulated lncRNAs, AX747207 was prominent because its associated gene RUNX3 was previously reported to be relative to malignant tumor chemoresistance. GO analysis results also indicated some biological processes and molecular functions linked to chemoresistance. The pathway enrichment results provided some potential pathways associated with chemoresistance. In the present study, the authors intended to identify lncRNA expression character in drug resistant cell line MCF‑7/ADR, corresponding to the parental MCF‑7 cell line. In addition, the study identified the lncRNA AX747207, and its potential targeted gene RUNX3, may be related to chemoresistance in breast cancer. These results may new insights into exploring the mechanisms of chemoresistance in breast cancer.
Insights
This study identifies long non-coding RNA (lncRNA) expression changes in doxorubicin-resistant breast cancer cells. The lncRNA AX747207 and its gene RUNX3 show potential links to chemoresistance mechanisms.
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- Breast cancer is a leading cause of cancer death globally, with chemoresistance significantly hindering treatment efficacy.
- Understanding the molecular mechanisms underlying chemoresistance is crucial for developing more effective therapeutic strategies.
Purpose of the Study:
- To identify long non-coding RNA (lncRNA) expression profiles in a doxorubicin-resistant breast cancer cell line (MCF-7/ADR) compared to its parental counterpart (MCF-7).
- To investigate the potential role of specific differentially expressed lncRNAs, such as AX747207, and their associated genes (e.g., RUNX3) in breast cancer chemoresistance.
Main Methods:
- Microarray analysis was performed to compare lncRNA expression between MCF-7 and MCF-7/ADR cells.
- Doxorubicin resistance was confirmed using MTT assay.
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to validate microarray findings. Gene Ontology (GO) and pathway analyses were conducted.
Main Results:
- A total of 8,892 differentially expressed lncRNAs were identified in MCF-7/ADR cells compared to MCF-7 cells (absolute fold-change >2.0).
- 32 lncRNAs were selected for validation via RT-qPCR.
- The lncRNA AX747207 was highlighted due to its association with RUNX3, a gene previously linked to chemoresistance in malignant tumors. GO and pathway analyses suggested biological processes and pathways relevant to chemoresistance.
Conclusions:
- The study successfully characterized lncRNA expression differences in a drug-resistant breast cancer cell line.
- lncRNA AX747207 and its potential target gene RUNX3 may play a significant role in breast cancer chemoresistance.
- These findings offer new insights into the molecular mechanisms of chemoresistance and potential therapeutic targets.
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