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Knockdown of long non‑coding RNA PVT1 reverses multidrug resistance in colorectal cancer cells
Heng Fan1, Jian-Hua Zhu2, Xue-Qing Yao1
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Abstract:
Multidrug resistance (MDR) is one of the primary causes of chemotherapy failure in colorectal cancer (CRC), and extensive biological studies into MDR are required. The non‑coding RNA plasmacytoma variant translocation 1 (PVT1) has been demonstrated to be associated with low survival rates in patients with CRC. However, whether PVT1 serves a critical function in the MDR of CRC remains to be determined. To determine the association between PVT1 expression and 5‑fluorouracil (5‑FU) resistance in CRC, the expression levels of PVT1 mRNA in 5‑FU‑resistant CRC tissues and cell lines (HCT‑8/5‑FU and HCT‑116/5‑FU) were assessed by a reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Cytotoxicity was evaluated using a Cell Counting Kit‑8 assay and apoptosis rates were assessed via flow cytometry. In the present study, PVT1 mRNA was highly expressed in 5‑FU‑resistant CRC tissues and cell lines. HCT‑8/5‑FU and HCT‑116/5‑FU cells transfected with small interfering RNA PVT1 and treated with 5‑FU exhibited higher apoptotic rates and lower survival rates. By contrast, overexpression of PVT1 in HCT‑8 and HCT‑116 cells transfected with lentiviral vector‑PVT1‑green fluorescent protein and treated with 5‑FU exhibited lower apoptosis rates and higher survival rates. RT‑qPCR and western blotting demonstrated that the overexpression of PVT1 increased the mRNA and protein expression levels of multidrug resistance‑associated protein 1, P‑glycoprotein, serine/threonine‑protein kinase mTOR and apoptosis regulator Bcl2. The present study indicates that PVT1 overexpression may promote MDR in CRC cells, and suggested that inhibition of PVT1 expression may be an effective therapeutic strategy for reversing MDR in CRC.
Insights
Plasmacytoma variant translocation 1 (PVT1) is highly expressed in multidrug-resistant colorectal cancer (CRC). Inhibiting PVT1 may reverse 5-fluorouracil resistance in CRC, offering a potential therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Multidrug resistance (MDR) is a major challenge in colorectal cancer (CRC) chemotherapy.
- Plasmacytoma variant translocation 1 (PVT1) is linked to poor survival in CRC patients.
- The role of PVT1 in CRC MDR is not fully understood.
Purpose of the Study:
- To investigate the association between PVT1 expression and 5-fluorouracil (5-FU) resistance in colorectal cancer.
- To determine if PVT1 plays a critical role in mediating MDR in CRC cells.
Main Methods:
- Assessed PVT1 mRNA levels in 5-FU-resistant CRC tissues and cell lines using RT-qPCR.
- Evaluated cytotoxicity via Cell Counting Kit-8 assay and apoptosis rates using flow cytometry.
- Manipulated PVT1 expression (siRNA knockdown and overexpression) in CRC cell lines.
Main Results:
- PVT1 mRNA was significantly upregulated in 5-FU-resistant CRC tissues and cell lines.
- PVT1 knockdown increased apoptosis and decreased survival in 5-FU-treated CRC cells.
- PVT1 overexpression decreased apoptosis and increased survival in 5-FU-treated CRC cells, upregulating MDR-associated proteins (MRP1, P-gp) and mTOR/Bcl2.
Conclusions:
- PVT1 overexpression promotes MDR in colorectal cancer cells.
- Inhibition of PVT1 may serve as a therapeutic strategy to overcome 5-FU resistance in CRC.
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