Pachymic Acid Ameliorates Pulmonary Hypertension by Regulating Nrf2-Keap1-ARE Pathway.

OBJECTIVE: Pulmonary hypertension (PH) is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death. The purpose of this study was to investigate the mechanism by which pachymic acid (PA) pretreatment affects PH and pulmonary vascular remodeling in rats. METHODS: PH was induced via hypoxia exposure and administration of PA (5 mg/kg per day) in male Sprague-Dawley rats. Hemodynamic parameters were measured using a right ventricular floating catheter and pulmonary vascular morphometry was measured by hematoxylin-eosin (HE), α-SMA and Masson staining. MTT assays and EdU staining were used to detect cell proliferation, and apoptosis was analyzed by TUNEL staining. Western blotting and immunohistochemistry were used to detect the expression of proteins related to the Nrf2-Keap1-ARE pathway. RESULTS: PA significantly alleviated hypoxic PH and reversed right ventricular hypertrophy and pulmonary vascular remodeling. In addition, PA effectively inhibited proliferation and promoted apoptosis in hypoxia-induced pulmonary artery smooth muscle cells (PASMCs). Moreover, PA pretreatment inhibited the expression of peroxy-related factor (MDA) and promoted the expression of antioxidant-related factors (GSH-PX and SOD). Furthermore, hypoxia inhibited the Nrf2-Keap1-ARE signaling pathway, while PA effectively activated this pathway. Most importantly, addition of the Nrf2 inhibitor ML385 reversed the inhibitory effects of PA on ROS generation, proliferation, and apoptosis tolerance in hypoxia-induced PASMCs. CONCLUSION: Our study suggests that PA may reverse PH by regulating the Nrf2-Keap1-ARE signaling pathway.

[miR-520a-3p regulates the signaling pathway of cytokine secretion in cervical cancer].

OBJECTIVE: To investigate the molecular mechanism of miR-520a-3p regulating the secretion of cytokines in cervical cancer. METHODS: The matching between miR-520a-3p and the NF-κB complex subunit RELA was analyzed by TargetScanHuman, and then the luciferase reporting system was used to detect whether miR-520a-3p targeted the NF-κB complex subunit RELA. After cervical cancer HELA cells stimulated by LPS, in the overexpression group or the knockout group, miR-520a-3p mimics or inhibitor were mixed with transfection reagent and dripped into HELA cells. The expression levels of colony stimulating factor (GCSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 19 (IL-9), interleukin 10 (IL-10), interleukin 12 p40 (IL-12 p40), interleukin 12 p70 (IL-12 p70), interleukin 13 (IL-13), interleukin 17 (IL-17), interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), monocyte chemoattractant protein-5 (MCP-5), small inducible cytokine A5 (RANTES), tumor necrosis factor-alpha (TNFα) were detected by enzyme-linked immunosorbent assay in both the overexpression group and the knockout group. Each experiment was repeated three times. RESULTS: miR-520a-3p targeted the 3'UTR of RELA. After activation of the NF-κB signaling pathway by lipopolysaccharide (LPS), the protein expression levels of cytokines GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα secreted by HELA cells of cervical cancer were increased significantly (P＜0.05). In the overexpression group, the protein expression level of RELA of NF-κB complex subunit was decreased, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were decreased (P＜0.05). In the knockdown group, the protein expression level of RELA, a subunit of NF-κB complex, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were increased (P＜0.05). CONCLUSION: miR-520a-3p inhibits cytokine secretion in HELA cells of cervical cancer by targeting RELA, a key molecule in the NF-κB signaling pathway.

Bisphosphonates enhance EGFR-TKIs efficacy in advanced NSCLC patients with EGFR activating mutation: A retrospective study.

BACKGROUND: Bisphosphonates have exhibited anti-tumor activity in non-small cell lung cancer (NSCLC). We aimed to evaluate whether the combination of bisphosphonates with tyrosine kinase inhibitors of EGFR (EGFR-TKIs) could obtain a synergistic effect on advanced NSCLC patients with EGFR mutations. METHODS: Between January 2008 and October 2013, 114 advanced EGFR mutations NSCLC patients who received EGFR-TKIs as first-line therapy were recruited from two cancer centers. Patients were separated into EGFR-TKIs alone or EGFR-TKIs plus bisphosphonates (combination) group. Median progression free survival (mPFS), median overall survival (mOS) distributions and survival curves were analyzed. RESULTS: Among the 114 patients, 62 had bone metastases (19 patients treated with EGFR-TKIs, 43 patients treated with EGFR-TKIs + bisphosphonates). Median PFS and OS were significantly improved in combination group compared with EGFR-TKIs group (mPFS: 15.0 vs 7.3 months, P = 0.0017; mOS: 25.2 vs 10.4 months, P = 0.0015) in patients with bone metastases. Among the 71 patients (19 patients with bone metastases) treated with EGFR-TKIs alone, patients with bone metastases had poor survival prognosis (mPFS:7.3 vs 12.1 months, P = 0.0434; mOS:10.4 vs 22.0 months, P = 0.0036). The survival of patients with bone metastases who received EGFR-TKIs plus bisphosphonates therapy was non-inferior to patients without bone metastases treated with EGFR-TKIs alone (mPFS: 15.0 vs 12.1 months, p = 0.1871; mOS: 25.2 vs 22.0 months, p = 0.9798). CONCLUSIONS: Concomitant use of bisphosphonates and EGFR-TKIs improves therapeutic efficacy and brings survival benefits to NSCLC patients with EGFR mutation and bone metastases.

Rapid increase of serum neuron specific enolase level and tachyphylaxis of EGFR-tyrosine kinase inhibitor indicate small cell lung cancer transformation from EGFR positive lung adenocarcinoma?

We report the case of an 80-year-old male with relapsed EGFR exon 19 deletion lung adenocarcinoma treated with EGFR-tyrosine kinase inhibitor (TKI), but with poor response and rapid increase of serum neuron specific enolase (NSE). Repeat biopsy identified pathological transformation to small cell lung cancers (SCLC) retaining the same EGFR mutation. This case highlights routine serological testing of NSE may benefit for the lung adenocarcinoma patients resistant to TKIs.

Risk factors for early recurrence of HBV-related hepatocellular carcinoma meeting milan criteria after curative resection.

BACKGROUND: The prognosis of patients with hepatocellular carcinoma (HCC) after curative resection varies greatly. Few studies had investigated the risk factors for early recurrence (recurrence-free time ≤ 1 year) of hepatitis B virus (HBV)-related HCCs meeting Milan criteria. METHODS: A retrospective analysis was performed on the 224 patients with HCC meeting Milan criteria who underwent curative liver resection in our center between February 2007 and March 2012. The overall survival (OS) rate, recurrence-free survival (RFS) rate and risk factors for early recurrence were analyzed. RESULTS: After a median follow-up of 33.3 months, HCC reoccurred in 105 of 224 patients and 32 died during the period. The 1-, 3- and 5-year OS rates were 97.3%, 81.6% and 75.6% respectively, and the 1-, 3- and 5-year RFS rates were 73.2%, 53.7% and 41.6%. Cox regression showed alpha-fetoprotein (AFP) > 800 ng/ml (HR 2.538, 95% CI 1.464-4.401, P=0.001), multiple tumors (HR 2.286, 95% CI 1.123-4.246, P=0.009) and microvascular invasion (HR 2.518, 95% CI 1.475-4.298, P=0.001) to be associated with early recurrence (recurrence-free time ≤ 1-year) of HCC meeting Milan criteria. CONCLUSIONS: AFP > 800 ng/ml, multiple tumors and microvascular invasion are independent risk factors affecting early postoperative recurrence of HCC. In addition resection appears capable of replacing liver transplantation in some situations with safety and a better outcome.