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Biological effects of RNAi targeted inhibiting Tiam1 gene expression on cholangiocarcinoma cells
Wei Cheng1, Yaling Liu1, Zhi Zuo1
1Department of Hepatobiliary Surgery, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University Changsha, Hunan, China.
Objective:
To investigate the characteristics of Tiam1 gene expression in human cholangiocarcinoma tissues and benign bile duct tissues, and to analyze the correlations between Tiam1 gene expression and the degree of tumor differentiation, invasive and metastatic abilities. To explore the effect of targeted inhibiting Tiam1 gene expression on proliferation and migration activity of human cholangiocarcinoma cells.
Methods:
Expression of Tiam1 in 83 cases of cholangiocarcinoma tissues and 25 cases of benign bile tissues was detected using immunohistochemistry. The clinical data of patients with cholangiocarcinoma were collected. The correlations between Tiam1 gene expression and the clinicopathologic features in patients with cholangiocarcinoma were analyzed. The human cholangiocarcinoma RBE cells were divided into 3 groups. Cells in experimental group and control group were respectively transfected with Tiam1 shRNA lentiviral vectors and negative shRNA lentiviral control vectors. Cells in blank group received no treatment. Real-time PCR endogenesis was used to verify Tiam1 gene expression. Cell cycle experiments and MTT assay were used to measure cell proliferation activity. Transwell test was used to detect cell migration activity.
Results:
The negative rate Tiam1 protein expression in cholangiocarcinoma tissues was significantly higher than that in benign bile tissues (P<0.001). Tiam1 protein expression in cholangiocarcinoma tissues had correlations with cholangiocarcinoma differentiation degree, TNM stage and lymph node metastasis (P<0.05), and had no significant correlations with gender, age and distant metastasis (P>0.05). Real-time PCR detection indicated that Tiam1 expression of experimental group was significantly lower than that in control group and blank group (P<0.05), demonstrating that Tiam1 shRNA was effective on Tiam1 gene silencing in RBE cells. Cell cycle experiment showed that the percentage of S phase in cell cycle in experimental group was lower than that in control group and blank group (P<0.05), demonstrating that after the down-regulation of Tiam1 gene expression, the speed of cell proliferation was inhibited. MTT assay results showed that the total growth speed in experimental group was significantly lower than that in control group and blank group (P<0.05), indicating that the proliferation activity of cholangiocarcinoma cells was inhibited after targeted inhibition of Tiam1 gene expression. Transwell detection results showed that the metastasis rate in experimental group was significantly lower than that in control group and blank group (P<0.05), demonstrating that targeted inhibition of Tiam1 gene expression could significantly inhibit migration ability of RBE cells.
Conclusion:
Tiam1 expression significantly increased in cholangiocarcinoma tissues, and increased along with the degree of malignancy of cholangiocarcinoma. Targeted silencing Tiam1 expression could inhibit proliferation and migration activity of cholangiocarcinoma cells.
Insights
Tiam1 gene expression is elevated in cholangiocarcinoma and linked to malignancy. Inhibiting Tiam1 in cancer cells reduces their proliferation and migration, suggesting Tiam1 as a potential therapeutic target for cholangiocarcinoma.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Cholangiocarcinoma (CCA) is a challenging malignancy with limited treatment options.
- Tiam1 (T-cell lymphoma invasion and metastasis 1) is implicated in cancer progression.
- Understanding Tiam1's role in CCA is crucial for developing targeted therapies.
Purpose of the Study:
- To examine Tiam1 gene expression in human CCA tissues versus benign bile ducts.
- To correlate Tiam1 expression with CCA clinicopathological features, including differentiation and metastasis.
- To investigate the impact of Tiam1 gene silencing on CCA cell proliferation and migration.
Main Methods:
- Immunohistochemistry used to quantify Tiam1 protein in 83 CCA and 25 benign tissues.
- Correlation analysis between Tiam1 expression and clinicopathological variables.
- In vitro study using RBE cells with Tiam1 shRNA lentiviral vectors for gene silencing.
- Real-time PCR, cell cycle analysis, MTT assay, and Transwell assays to assess proliferation and migration.
Main Results:
- Tiam1 protein expression was significantly higher in CCA tissues than benign tissues (P<0.001).
- Tiam1 expression correlated with CCA differentiation, TNM stage, and lymph node metastasis (P<0.05).
- Tiam1 gene silencing in RBE cells significantly inhibited proliferation (cell cycle, MTT) and migration (Transwell) (P<0.05).
Conclusions:
- Tiam1 is upregulated in cholangiocarcinoma and associated with increased malignancy.
- Targeted inhibition of Tiam1 effectively suppresses CCA cell proliferation and migration.
- Tiam1 represents a promising therapeutic target for cholangiocarcinoma treatment.
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