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MiR-32-5p Regulates Radiosensitization, Migration And Invasion Of Colorectal Cancer Cells By Targeting TOB1 Gene
Hong Liang1, Yumei Tang1, Hui Zhang1
1Department of Gastrointestinal Surgery, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, School of Clinical Medicine, Henan University, Zhengzhou, Henan 450003, People's Republic of China.
Background:
MicroRNAs (miRNAs) play key roles in the development and progression of various cancers. However, the precise functions and regulation mechanisms of miRNAs in human tumors remain elusive.
Methods:
Quantitative real time-PCR (qRT-PCR) was performed to detect the level of miR-32-5p in colorectal cancer tissues. The relationships between miR-32-5p level with clinicopathological characteristics and prognosis were analyzed. The miR-32-5p inhibitor was employed to knock down the expression of miR-32-5p. The overexpression plasmid and si-RNA targeting TOB1 were generated. Clone formation assays under radiant exposure were used to evaluate the radiosensitization. Transwell migration and invasion were employed to test the ability of cell migration and invasion. Luciferase reporter assays were used to confirm the regulation of miR-32-5p on the expression of TOB1. Rescue experiments were conducted to investigate the effects of TOB1 on the functions of miR-32-5p.
Results:
In this study, we found that miR-32-5p was significantly upregulated in colorectal cancer tissues compared with adjacent normal tissues. The level of miR-32-5p was positively correlated with tumor differentiation and metastasis. Log-rank tests showed that high level of miR-32-5p was significantly correlated with poor overall survival and disease-free survival. Anti-miR-32-5p remarkably enhanced the radiosensitivity and inhibited migration and invasion of colorectal cancer cells. In addition, overexpression of TOB1 obviously increased the radiosensitivity and inhibited migration and invasion of colorectal cancer cells. Moreover, bioinformatics analysis and luciferase reporter assays demonstrated that miR-32-5p suppressed the expression of TOB1 through directly binding to the 3'-UTR of TOB1 mRNA. Rescue experiments indicated that miR-32-5p regulated the radiosensitivity, migration and invasion of colorectal cancer cells through inhibiting TOB1 expression.
Conclusion:
This study suggested that miR-32-5p may serve as a prognostic and therapeutic target for colorectal cancer, and downregulation of miR-32-5p enhanced the radiosensitivity and inhibited migration and invasion through promoting TOB1 expression.
Insights
MicroRNA-32-5p (miR-32-5p) is upregulated in colorectal cancer, correlating with poor prognosis. Downregulating miR-32-5p enhances radiosensitivity and inhibits cancer cell migration and invasion by increasing TOB1 expression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are crucial in cancer development, but their specific roles in human tumors are not fully understood.
- Investigating novel miRNA targets is essential for advancing cancer therapy.
Purpose of the Study:
- To elucidate the role of miR-32-5p in colorectal cancer.
- To determine the relationship between miR-32-5p, radiosensitivity, and metastasis.
- To identify therapeutic targets for colorectal cancer treatment.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to measure miR-32-5p levels in colorectal cancer tissues.
- In vitro assays including clone formation, Transwell migration/invasion, and luciferase reporter assays.
- Manipulation of miR-32-5p and TOB1 expression using inhibitors, overexpression plasmids, and si-RNA.
Main Results:
- miR-32-5p was significantly upregulated in colorectal cancer tissues and correlated with poor survival.
- Downregulation of miR-32-5p increased radiosensitivity and reduced migration and invasion.
- miR-32-5p directly suppressed TOB1 expression, and TOB1 overexpression mimicked the effects of miR-32-5p inhibition.
Conclusions:
- miR-32-5p is a potential prognostic biomarker and therapeutic target for colorectal cancer.
- Targeting miR-32-5p may enhance radiosensitivity and inhibit colorectal cancer progression by modulating TOB1.
- Downregulation of miR-32-5p represents a promising strategy for colorectal cancer treatment.
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