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[Triptolide inhibits cell proliferation by downregulating phosphorylation of estrogen reporters in 4T1 tumor-bearing
Abstract:
In order to investigate the anti-proliferative effects of triptolide (TP) on 4T1 mice breast cancer cell line in vitro and in mouse model, as well as the possible mechanisms, we detected the effect of TP on cell proliferation by MTT assay or Crystal Violet Staining in our research. Flowcytometry combined with FITC-Annexin V/PI staining were used for detecting TP induced 4T1 cell apoptosis. The protein expression of ERalpha, p-ERalpha, ERbeta, p-ERbeta, ERK, p-ERK, p38, p-p38, SAPK/JNK, and p-SAPK/JNK was tested by western blotting. We also compare TP with chemotherapy drug doxorubicin in 4T1 tumor bearing BLAB/c mice model, the Xenogen bioluminescence imaging, H&E, and IHC result indicated that TP exhibits an anticancer proliferation activity. As a result, TP in 100, 10, 1, 0.1 micromol x L(-1), all inhibited the proliferation of 4T1 cells by MTT assay and Crystal Violet Staining. TP which concentrations is 10, 1, 0.1 micromol x L(-1) could induce the apoptosis of 4T1 cells and reduce the cell proliferation. TP in 200 microg x kg(-1) could inhibit the tumor growth in vivo. The anticancer proliferation of TP was involved in its effect on reducing expression of ERalpha, p-ERalpha, ERbeta, and p-ERbeta, but nothing to do with the activation of MAPK signaling pathway.
Insights
Triptolide (TP) effectively inhibits 4T1 breast cancer cell proliferation and induces apoptosis in vitro and in vivo. Its anti-cancer effects are linked to reduced estrogen receptor expression, not MAPK pathway activation.
Area of Science:
- Pharmacology
- Oncology
- Molecular Biology
Context:
- Breast cancer remains a significant health concern, necessitating novel therapeutic strategies.
- 4T1 murine breast cancer cell line is a widely used model for preclinical cancer research.
- Triptolide (TP), a compound from Tripterygium wilfordii, has shown potential anti-cancer properties.
Purpose:
- To investigate the anti-proliferative and apoptosis-inducing effects of triptolide (TP) on the 4T1 breast cancer cell line.
- To elucidate the underlying molecular mechanisms, including the role of estrogen receptors and MAPK signaling pathways.
- To compare the efficacy of TP with doxorubicin in a 4T1 tumor-bearing mouse model.
Summary:
- TP significantly inhibited 4T1 cell proliferation in vitro across various concentrations (0.1-100 µmol/L) as assessed by MTT assay and Crystal Violet Staining.
- TP induced apoptosis in 4T1 cells at concentrations of 0.1-10 µmol/L, leading to reduced cell proliferation.
- In vivo, TP (200 µg/kg) demonstrated anti-tumor activity in a 4T1 tumor-bearing mouse model, evidenced by bioluminescence imaging, H&E, and IHC.
- TP's anti-cancer effects were associated with decreased expression of estrogen receptors (ERalpha, p-ERalpha, ERbeta, p-ERbeta), but not with the activation of the MAPK signaling pathway.
Impact:
- This study highlights triptolide's potential as a therapeutic agent for breast cancer.
- Understanding TP's mechanism of action, particularly its interaction with estrogen receptors, can guide future drug development.
- The findings provide a basis for further investigation into TP's clinical applicability in breast cancer treatment.
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