Anti-Cervical Cancer Role of Matrine, Oxymatrine and Sophora Flavescens Alkaloid Gels and its Mechanism

Abstract

Background: Cervical cancer is one of the leading severe malignancies throughout the world. Sophra flavescens alkaloid (SFA) gels, a compound Traditional Chinese Medicine, has been clinically used in China for many years. Its individual active ingredients are matrine and oxymatrine, which has been showed that they can restrain primary tumorigenesis, while the underlying molecular mechanisms of SFA gels in cervical cancer cells remain unclear. Methods: To detect the effect of SFA gels and its active ingredients, CCK-8 assay and colony assay were used on cervical cancer cells proliferation. Transwell assay was used to detect cancer cell migration. Apoptosis and cell cycle arrest were used to detect whether SFA gels effect the cervical cancer cells proliferation. Western blot was used to detect whether SFA gels regulate the cervical cancer cells via the suppression of AKT/mTOR signaling pathway. Results: SFA gels can restrain cervical cancer cell proliferation, inhibit metastasis, induce cell cycle arrest in G2/M phase, induce cellular apoptosis through stimulation of Bax and E-cadherin, and suppression of Bcl-2, cyclin A, MMP2. Further study shows that SFA gels may regulate the cervical cancer cells via the suppression of AKT/mTOR signaling pathway. Conclusions: SFA gels, like its active ingredients, can restrain cervical cancer cells proliferation, suppress cervical cancer cell migration, induce the apoptosis and cell cycle arrest in cervical cancer cells. SFA gels may be a potential anti-tumor therapeutic agent for treating cervical cancer.

Keywords: AKT; Cervical cancer; Matrine; Oxymatrine; Sophora flavescens alkaloid gels; mTOR.

Figure 1

Sophora flavescens alkaloid gels and its active ingredients can restrain cervical cells proliferation. (A), (B) and (C) Growth inhibition assay of SiHa, C33A, Caski and HK-2 was detected by CCK8 assay under different concentrations of Sophora flavescens alkaloid gels (A), matrine (B) and oxymatrine (C) for 48 hours. (D) colony assay of SiHa was used to further measure Sophora flavescens alkaloid gels' growth inhibitory effect.

Figure 2

Sophora flavescens alkaloid gels suppress cervical cancer cell migration. (A), (C) Cell migration was detected by the transwell assay. (B), (D) Columns represent quantification of invaded cells, *P < 0.05, **P<0.01. Error bars = 95% Cis. (E) Expression of cell adhesion-associated protein E-cadherin and MMP2 were detected by Western Blot.

Figure 3

Sophora flavescens alkaloid gels induces the apoptosis and cell cycle arrest in cervical cancer cells. (A) Flow cytometry apoptosis analysis was performed in cultured Siha, C33A and Ca ski cell lines harvested after 2 days of treatment with different concentration of SFA gels, matrine and oxymatrine. (B) Quantification of apoptosis for the cell lines. *P<0.05, **P<0.01. Columns are means of three independent experiments, bars are SD. (C) Cell cycle analysis was performed using flow cytometry by staining with propidium iodide. (D) Columns represent quantification of cell phase. (E) and (F) Western Blot analysis of apoptosis-associated and cell cycle regulatory proteins.

Figure 4

Sophora flavescens alkaloid gels inhibit the cervical cancer cell anti-cancer activities through AKT/mTOR signaling pathways. Expression of the components of the AKT/mTOR pathways were detected by Western blot in both SiHa and C33A cell lines.