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The anti-cancerous activity of recombinant trichosanthin on prostate cancer cell PC3
JinLong Li1,2, Hui Li3, ZhaoLi Zhang4
1Department of Laboratory Medicine, The Second Affiliated Hospital of Southeast University, Zhongfu Road 1-1, Nanjing, 210003, China.
Context:
Trichosanthin produced in the root tube of Trichosanthes kirilowii shows anti-tumor activity on a series of cancer cells including Hela, MCF-7, HL-60. But there is little information about its effect on the carcinogenesis of prostate cancer.
Objective:
This work was designed to study the role of trichosanthin on prostate cancer cells PC3.
Materials And Methods:
Trichosanthin was expressed in BL21 strain and purified by affinity chromatography. MTT assay was designed to determine the effect of trichosanthin on growth of PC3 cells at doses of 10, 20, 40, 60, 80, and 120 μg/ml. Then the effect of 50 μg/ml rTCS alone or combined with 2 μM IL-2 on PC3 cell proliferation was analyzed. And the mechanism of rTCS was studied by western blot. After that the in vivo effect of rTCS combined with IL-2 was explored in mice bearing PC3 xenograft tumor.
Results:
Trichosanthin was successfully expressed in BL21 and purified by 100 mM imidazole. It was shown to inhibit proliferation of PC3 cells in a dose-dependent manner with IC50 50.6 μg/ml. When combined with cytokine IL-2, a significant synergic effect was obtained. The inhibition rate on PC3 was around 50 % in combination group while only 35.5 % in single rTCS group at 50 μg/ml. Further, the expression of full length caspase-8 and Bcl-2 decreased significantly while cleaved caspase-8 and Bax were up-regulated, which suggest that caspase-8-mediated apoptosis pathway may be activated by rTCS in PC3 cells. Moreover, our data demonstrated that tumor volume and tumor weight were significantly reduced in rTCS-treated or rTCS/IL-2-treated nude mice bearing PC3 xenograft tumor compared with control. And significant difference was also found between rTCS and rTCS/IL-2 group.
Conclusions:
This study demonstrates that rTCS is a potential agent with high in vitro and in vivo anti-tumor activity on PC3 cells. And rTCS combined with IL-2 is a promising strategy in treating patients with prostate cancer in future.
Insights
This study shows trichosanthin (rTCS) effectively inhibits prostate cancer cell growth in vitro and in vivo. Combining rTCS with IL-2 demonstrates a promising synergistic anti-tumor strategy for prostate cancer treatment.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Trichosanthin (TCS) exhibits anti-tumor properties against various cancer cells.
- Limited research exists on TCS's impact on prostate cancer development.
- This study investigates TCS's role in PC3 prostate cancer cells.
Purpose of the Study:
- To evaluate the anti-prostate cancer effects of trichosanthin (rTCS) on PC3 cells.
- To explore the synergistic potential of rTCS combined with IL-2.
- To elucidate the underlying molecular mechanisms of rTCS-induced apoptosis.
Main Methods:
- Recombinant trichosanthin (rTCS) was expressed and purified.
- MTT assays assessed rTCS's effect on PC3 cell proliferation.
- Western blotting analyzed apoptosis-related protein expression.
- In vivo studies utilized mice bearing PC3 xenograft tumors.
Main Results:
- rTCS inhibited PC3 cell proliferation dose-dependently (IC50 = 50.6 μg/ml).
- Combination therapy with IL-2 showed significant synergistic anti-proliferative effects.
- rTCS induced apoptosis via the caspase-8 pathway, indicated by altered protein levels (caspase-8, Bcl-2, Bax).
- rTCS and rTCS/IL-2 significantly reduced tumor volume and weight in vivo.
Conclusions:
- rTCS demonstrates potent in vitro and in vivo anti-tumor activity against PC3 prostate cancer cells.
- The combination of rTCS and IL-2 presents a promising therapeutic strategy for prostate cancer.
- rTCS activates caspase-8-mediated apoptosis, contributing to its anti-cancer effects.
