Effects of lentiviral-mediated Foxp1 and Foxq1 RNAi on the hepatocarcinoma cell
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Updated: May 6, 2026
Cell Type-specific Gene Expression Profiling in the Mouse Liver
Published on: September 17, 2019
Jing Qin1, Yuyin Xu2, Xingyu Li1
1Department of Pathological Anatomy, Nantong University, Nantong, China.
Abstract:
Foxp1 and Foxq1 are two multifunctional molecules of "forkhead box (Fox)" family. The objective of this paper was to construct the lentiviral vectors expressing RNA interference (RNAi) against Foxp1 or Foxq1 genes, and the effects of both vectors with two RNAis on the proliferation, migration and apoptosis of 7721 hepatocarcinoma cell line were evaluated. Six target sequences against human Foxp1/Foxq1 mRNA were designed respectively and six pairs of their corresponding double-strand DNA oligo (siRNA) were synthesized prior to being transfected into 7721 cells with lipo2000, then a most efficient siRNA were selected to be subcloned into pLL3.7-GFP/Lenti plasmids. These plasmids were transfected into 293T cells to package lentiviral particles for subsequent transfection into 7721 cells after their sequences were confirmed. The expression of Foxp1and Foxq1 genes in the transfected cells were identified by real-time PCR. The migration, infiltration, viability and apoptosis of the transfected cells were assessed by wound healing assay, Transwell assay, CCK-8 assay and flow cytometry. Sequencing results showed that lentiviral vectors contained Foxp1 or Foxq1 gene. After being transfected into 7721 cells, Foxp1 and Foxq1 expression were significantly down-regulated by siRNA-823 and siRNA-834. The migration and infiltration ability, and the viability of 7721 cells transfected with two siRNAs were significantly suppressed; flow cytometry assay exhibited the apoptosis rate of transfected 7721 cells with the lentivirus RNAi vector of Foxp1 or Foxq1 was increased. All the results showed that the lentivirus RNAi vectors of Foxp1 and Foxq1 were able to inhibit the expression of Foxp1 and Foxq1 in 7721 cells efficiently, and the down-regulation of either Foxp1 or Foxq1 resulted in suppression of migration, infiltration and viability of 7721 cells and an increase in cell apoptosis. Our data indicated that both Foxp1 and Foxq1 genes played an oncogenic role in hepatocarcinoma cells, which proposed the two genes as new therapeutic targets for the cancer.
Insights
Forkhead box P1 (Foxp1) and Forkhead box Q1 (Foxq1) genes promote hepatocarcinoma cell growth and spread. Inhibiting these genes with lentiviral RNA interference suppressed tumor cell proliferation, migration, and invasion, while increasing apoptosis.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Forkhead box (Fox) family proteins, including Foxp1 and Foxq1, are implicated in various cellular processes.
- Hepatocellular carcinoma (HCC) is a major global health concern with limited therapeutic options.
- Understanding the role of specific genes in HCC progression is crucial for developing targeted therapies.
Purpose of the Study:
- To construct lentiviral vectors for RNA interference (RNAi) targeting Foxp1 and Foxq1 genes.
- To evaluate the effects of Foxp1 and Foxq1 knockdown on the proliferation, migration, and apoptosis of the 7721 hepatocarcinoma cell line.
- To investigate the potential of Foxp1 and Foxq1 as therapeutic targets in HCC.
Main Methods:
- Design and synthesis of six short interfering RNA (siRNA) sequences targeting human Foxp1/Foxq1 mRNA.
- Selection of the most efficient siRNA and subcloning into lentiviral vectors (pLL3.7-GFP/Lenti).
- Lentiviral particle production and transfection into 7721 hepatocarcinoma cells; assessment of gene expression, cell proliferation, migration, invasion, and apoptosis using real-time PCR, CCK-8, wound healing, Transwell, and flow cytometry assays.
Main Results:
- Lentiviral vectors successfully expressed shRNAs targeting Foxp1 and Foxq1, leading to significant gene down-regulation in 7721 cells.
- Knockdown of Foxp1 and Foxq1 significantly suppressed cell proliferation, migration, and invasion.
- Increased apoptosis was observed in 7721 cells following Foxp1 or Foxq1 gene silencing.
Conclusions:
- Lentiviral RNAi vectors effectively inhibit Foxp1 and Foxq1 expression in hepatocarcinoma cells.
- Down-regulation of Foxp1 or Foxq1 suppresses key oncogenic properties of HCC cells, including proliferation, migration, and invasion.
- Foxp1 and Foxq1 play significant oncogenic roles in HCC and represent promising therapeutic targets for cancer treatment.
