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[Construction of a lentivirus interfering vector targeting Cyc1 and its interfering efficiency in nasopharyngeal
Zhen Liu1, Hong-yan Wang, Jie Long
1Department of Pathology, Guangzhou Medical College, China. narcissus_jane@163.com
Objective:
To analyze the expression of Cyc1 in nasopharyngeal carcinoma (NPC) and evaluate the interfering efficiency of a lentivirus interfering vector targeting Cyc1 in NPC cells.
Methods:
Microarray technique was used to examine the expression of Cyc1 in NPC tissues. Real-time PCR was utilized to confirm the high expression of Cyc1 in NPC tissues and NPC cell lines. The recombinant Cyc1 shRNA-expressing plasmid (pLentiU6/Cyc1-shRNA) was stably transfected into NPC cells, and the interfering efficiency against Cyc1 was evaluated by quantitative RT-PCR.
Results:
The result of microarray showed that Cyc1 was highly expressed in NPC tissues compared to noncancerous nasopharyngeal tissues, as confirmed by Real-time PCR. All of the 8 NPC cells showed a high expression of Cyc1, among which 5-8F cells showed the highest expression. Sequence analysis indicated that the recombinant plasmid pLentiU6/Cyc1-shRNA was successfully constructed and could significantly and stably suppress the expression of Cyc1 in NPC cells.
Conclusion:
Cyc1 is highly expressed in NPC cells. The lentivirus vector constructed can markedly inhibit the expression of Cyc1 in NPC cells, which provides assistance in the investigation of the function and molecular mechanism of Cyc1 in NPC.
Insights
Cyclin 1 (Cyc1) is highly expressed in nasopharyngeal carcinoma (NPC) cells. A lentivirus vector effectively suppressed Cyc1 expression in NPC cells, aiding further research into its role in NPC.
Area of Science:
- Oncology
- Molecular Biology
- Gene Expression Analysis
Background:
- Nasopharyngeal carcinoma (NPC) is a significant health concern.
- The role of specific genes, such as Cyclin 1 (Cyc1), in NPC pathogenesis requires further elucidation.
Purpose of the Study:
- To investigate the expression levels of Cyc1 in NPC tissues and cell lines.
- To assess the efficacy of a lentivirus-based short hairpin RNA (shRNA) vector in suppressing Cyc1 expression within NPC cells.
Main Methods:
- Microarray analysis and quantitative Real-time Polymerase Chain Reaction (RT-PCR) were employed to determine Cyc1 expression.
- Stable transfection of NPC cells with a recombinant Cyc1 shRNA-expressing lentivirus vector (pLentiU6/Cyc1-shRNA).
- Quantitative RT-PCR was used to evaluate the interfering efficiency of the lentivirus vector.
Main Results:
- Cyc1 was found to be significantly upregulated in NPC tissues compared to non-cancerous nasopharyngeal tissues.
- All tested NPC cell lines exhibited high Cyc1 expression, with the 5-8F cell line showing the highest levels.
- The constructed pLentiU6/Cyc1-shRNA vector successfully and stably inhibited Cyc1 expression in NPC cells.
Conclusions:
- Cyc1 is demonstrably overexpressed in NPC cells.
- The developed lentivirus vector provides a potent tool for inhibiting Cyc1 expression in NPC.
- This study facilitates future investigations into the functional role and molecular mechanisms of Cyc1 in NPC development.
