Telocinobufagin, a PLK1 suppressor that inhibits tumor growth and metastasis by modulating CDC25c and CTCF in HNSCC cells.

BACKGROUND: The high metastasis and mortality rates of head and neck squamous cell carcinoma (HNSCC) urgently require new treatment targets and drugs. A steroidal component of ChanSu, telocinobufagin (TBG), was verified to have anti-cancer effects in various tumors, but its activity and mechanism in anti-HNSCC were still unknown. PURPOSE: This study tried to demonstrate the anti-tumor effect of TBG on HNSCC and verify its potential mechanism. METHODS: The effect of TBG on cell proliferation and metastasis were performed and the TBG changed genes were detected by RNA-seq analysis in HNSCC cells. The GSEA and PPI analysis were used to identify the pathways targeted for TBG-regulated genes. Meanwhile, the mechanism of TBG on anti-proliferative and anti-metastasis were investigated in vitro and in vivo. RESULTS: The in vitro and in vivo experiments confirmed that TBG has favorable anti-tumor effects by induced G2/M phase arrest and suppressed metastasis in HNSCC cells. Further RNA-seq analysis demonstrated the genes regulated by TBG were enriched at the G2/M checkpoint and PLK1 signaling pathway. Then, the bioinformatic analysis of clinical data found that high expressed PLK1 were closely associated with poor overall survival in HNSCC patients. Furthermore, PLK1 directly and indirectly modulated G2/M phase and metastasis (by regulated CTCF) in HNSCC cells, simultaneously. TBG significantly inhibited the protein levels of PLK1 in both phosphorylated and non-phosphorylated forms and then, in one way, inactivated PLK1 failed to activate G2/M phase-related proteins (including CDK1, CDC25c, and cyclin B1). In another way, be inhibited PLK1 unable promote the nuclear translocation of CTCF and thus suppressed HNSC cell metastasis. In contrast, the anti-proliferative and anti-metastasis effects of TBG on HNSCC cell were vanished when cells high-expressed PLK1. CONCLUSION: The present study verified that PLK1 mediated TBG induced anti-tumor effect by modulated G2/M phase and metastasis in HNSCC cells.

[Effect of Jian'erle Granule on Thl7/Treg Imbalance of Asthma Mice].

Objective To observe the levels of Th17 associated inflammatory factor IL-17 and Treg associated inflammatory factor IL-10 in serum and bronchoalveolar lavage fluid (BALF) of bronchial asthma mice, and to explore the mechanism of Jian'erle Granule (JG) for preventing and treating asth- ma. Methods Totally 40 Balb/c mice were divided into 4 groups according to random digit table, i.e., the normal control group, the asthma group, the post-excitation asthma group, the JG +asthma group, 10 in each group. Asthma model was established by 1% ovalbumin (OVA) (grade V) solution sensitization and excitation after intraperitoneal injection of antigen suspension in all groups except the normal control group. JG (0. 36 g/mL) was administrated to mice in the JG +asthma group after modeling. Equal volume of normal saline was administrated to mice in the rest 3 groups. All medication lasted for 14 successive days. After medication mice in the JG +asthma group and the asthma group were excited with 1% OVA (grade I) again for 40 min by atomization inhalation. Lung inflammation was examined by HE staining. Levels of IL-10 and IL-17 in BALF and serum were measured by ELISA. Results Bronchial structure in lung tissue was normal in mice of the normal control group, with no inflammatory infiltration seen. Mucosal epithelial cells of bronchial wall were injured in the asthma group and the post-excitation asthma group, with inflammatory infiltration seen. Inflammation around bronchus and blood vessels was obviously attenuated in the JG+asthma group. Compared with the normal control group,the IL-17 level in serum and BALF significantly increased, IL-10 level significantly decreased in the asthma group (P <0.01). Compared with the asthma group, IL-17 level in serum and BALF increased and IL-10 level in serum and BALF decreased in the post-excitation asthma group (P <0.01). Compared with the asthma group, the IL-17 level in serum and BALF significantly decreased, IL-10 level significantly increased in the JG +asth- ma group (P <0. 01). Compared with the post-excitation.asthma group,the IL-17 level in serun and BALF significantly decreased and IL-10 level in serum and BALF significantly increased in the JG + asthma group (P <0. 01). Conclusion The mechanism for JG preventing and treating asthma might be correla- ted with regulating Th17/Treg cytokine balance in serum and BALF.