Related Experiment Video
Updated: Jan 16, 2026
Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
Published on: February 8, 2018
The concurrent silencing of Tim-3 and STAT-3 promotes tumor regression both in vitro and in ovo
Reza Karami1,2, Shahla Khodayari1, Farzaneh Eshaghi1
1Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Background:
Due to the intricate nature of the tumor microenvironment and the impairment of the anti-tumor responses of the immune system, finding a novel approach in the field of immunology-based therapy might influence the prognosis and survival of patients suffering from cancer. T-cell immunoglobulin and mucin-domain containing-3 (Tim-3) is a regulatory element of immune surveillance that exerts a pivotal function in the microenvironment of tumors. Although the precise functions of the Tim-3 remain incompletely understood, it has been established that it not only contributes to T cell exhaustion but also participates in the STAT-3/NF-κB (signal transducer and activator of transcription 3 / nuclear factor-κB) pathway, which plays a major role in the progression of tumors.
Methods:
In this research, we assessed the efficacy of co-blocking the Tim-3 and STAT-3 factors in inhibiting cancer cell growth. Therefore, we suppressed the expression of these factors in murine-derived malignant cell lines (4T1 and CT26), using siRNA (small interfering RNA) molecules encapsulated in chitosan lactate-based nano carriers we previously developed.
Results:
Transfecting the siRNAs into cancer cells with nanocarriers significantly downregulated the expression of Tim-3 and STAT-3 in both 4T1 and CT26 cells. Downregulation of Tim-3 and STAT-3 was correlated with diminished viability, proliferation, angiogenesis, and metastatic characteristics of cancerous cells, in vitro. Furthermore, co-silencing of Tim-3 and STAT-3 led to tumor regression, in ovo.
Conclusion:
These results revealed that the concurrent silencing of Tim-3 and STAT-3 can significantly suppresses the tumor growth by reducing cell proliferation, angiogenesis and metastasis in vitro and in ovo. However, future investigations-particularly in vivo models-are necessary to validate the current strategy as a potential anti-tumor therapeutic approach.
Insights
Co-blocking T-cell immunoglobulin and mucin-domain containing-3 (Tim-3) and signal transducer and activator of transcription 3 (STAT-3) using siRNA nanocarriers inhibited cancer cell growth. This novel immunology-based therapy approach suppressed tumor progression in vitro and in ovo.
Area of Science:
- Immunology
- Cancer Biology
- Nanotechnology
Background:
- The tumor microenvironment and impaired immune responses hinder cancer therapy.
- T-cell immunoglobulin and mucin-domain containing-3 (Tim-3) is crucial in tumor immune evasion and T-cell exhaustion.
- Tim-3 and STAT-3 pathways are implicated in tumor progression.
Purpose of the Study:
- To investigate the efficacy of co-blocking Tim-3 and STAT-3 for cancer treatment.
- To assess the potential of siRNA-loaded nanocarriers for delivering therapeutic agents.
Main Methods:
- Developed chitosan lactate-based nanocarriers for siRNA delivery.
- Suppressed Tim-3 and STAT-3 expression in murine cancer cell lines (4T1, CT26) using siRNA.
- Evaluated the effects of co-silencing on cancer cell characteristics and tumor growth.
Main Results:
- Nanocarrier-mediated siRNA transfection effectively downregulated Tim-3 and STAT-3 expression.
- Co-silencing correlated with reduced cancer cell viability, proliferation, angiogenesis, and metastasis in vitro.
- Tumor regression was observed in ovo following concurrent Tim-3 and STAT-3 silencing.
Conclusions:
- Concurrent silencing of Tim-3 and STAT-3 shows significant anti-tumor potential by inhibiting proliferation, angiogenesis, and metastasis.
- This strategy warrants further investigation in in vivo models as a potential cancer therapeutic approach.
