HSP70 promotes pancreatic cancer cell epithelial-mesenchymal transformation and growth via the NF-κB signaling pathway.

OBJECTIVE: To study the effects of HSP70 on proliferation, migration, invasion, and epithelial-mesenchymal transformation (EMT) of pancreatic cancer cells and explore its underlying mechanisms. METHODS: Pancreatic cancer cell models with either reduced HSP70 or increased HSP70 expression were established. RT-qPCR and Western blot assays were used to determine mRNA and protein levels of HSP70, IKK/IκBa/NF-κB signaling pathway-related genes, and EMT markers. The CCK-8 and cell cloning assays were used to evaluate cell proliferation and cloning abilities. Transwell and wound healing assays were used to assess the invasive and migratory properties of the cells. Effects of NF-κB signaling modulation were explored using an IKK inhibitor (BAY11-7082) and an IKK overexpression vector (pCMV-IKK). Electrophoretic mobility shift assay (EMSA) and luciferase reporter assays were conducted to analyze NF-κB's promoter binding and transcriptional activities. RESULTS: HSP70 knockdown inhibited p-p65 nuclear translocation and reduced the expression of p-p65, p-IKKα/ß, p-IκBα, N-cadherin, Vimentin, and Twist. It also decreased NF-κB's promoter binding and transcriptional activities, increased E-cadherin levels, and suppressed pancreatic cancer cell proliferation, cloning, migration, and invasion. In contrast, HSP70 overexpression led to increased expression of p-p65, p-IKKα/ß, p-IκBα, N-cadherin, Vimentin, and Twist, decreased E-cadherin levels, and enhanced cell proliferation, cloning, migration, and invasion capabilities. NF-κB signaling pathway modulation reversed EMT changes induced by altered HSP70 expression levels. rhHSP70 also increased p-IKKα/ß and p-IκBα protein levels. CONCLUSION: HSP70 promotes the EMT and enhances pancreatic cancer cell proliferation, migration, and invasion by activating the NF-κB pathway.

Serum HSP70 and VEGF Levels Are Effective Predictive Factors of Chemoradiosensitivity and Prognosis of Pancreatic Cancer Patients.

AIM: The study is to evaluate serum HSP70 and VEGF for predicting the chemoradiosensitivity of the pancreatic cancer patients. MATERIALS AND METHODS: 255 pancreatic cancer patients and 60 healthy subjects were measured for serum HSP70 and VEGF using ELISA for the pretreatment, during treatment, and postchemoradiotherapy timepoints. RESULTS: The serum HSP70 and VEGF were found to be elevated in pancreatic cancer patients as compared to healthy subjects. After chemoradiotherapy treatment, 179 patients showed effective clinical response while 76 patients showed ineffective clinical response. Serum HSP70 and VEGF were higher during chemoradiotherapy, and lower posttreatment in the effective group. However, serum HSP70 and VEGF were higher during and after treatment in the ineffective group. At any given timepoint, serum HSP70 and VEGF were higher in the ineffective group compared with the effective group. The overall survival and progression-free survival trends were as follows: HSP70 High /VEGF High < HSP70 High /VEGF Low or HSP70 Low /VEGF High < HSP70 Low /VEGF Low . Serum HSP70 and VEGF were individually effective, and their combination was even more effective in predicting the chemoradiosensitivity of pancreatic cancer patients. HSP70 and VEGF were independent risk factors for overall survival and progression-free survival of pancreatic cancer patients. CONCLUSIONS: Low levels of serum HSP70 and VEGF were associated with improved radiosensitivity and better prognosis of pancreatic cancer patients.

MiR-12200-5p Targets Multiple Members of Wnt Signaling Pathway to Inhibit Osteoblast Differentiation and Bone Formation.

BACKGROUND: Osteoporosis is widespread and has become an emerging problem in the elderly. MicroRNAs could affect osteoblast differentiation and further regulate the occurrence of osteoporosis by targeting osteogenic differentiation signaling pathways. Our screening study found that miR-12200-5p simultaneously targeted six important factors within the Wnt signaling pathway (Apc, Tcf4, Tcf7, Wnt3a, Wnt5a, and Lrp6), indicating that miR-12200-5p might function as a strong regulator of this pathway. Since the Wnt pathway exists as one of the most essential pathways for osteogenic differentiation, miR-12200-5p may have an important role in the development of osteoporosis. OBJECTIVE: This study intended to explore the regulatory role and corresponding mechanism of miR-12200-5p in osteoblast differentiation. METHODS: We investigated the differentiation of osteoblast after the treatments of miR-12200-5p mimic and inhibitor. The interactions between miR-12200-5p and its target genes were also detected. Furthermore, the rescue effect of miR-12200-5p inhibitor on osteoporosis was evaluated using an ovariectomized osteoporosis mouse model. RESULTS: MiR-12200-5p significantly inhibited osteoblast differentiation, and bound with the 3'-UTR sequences of its target genes (Apc, Tcf4, Tcf7, Wnt3a, Wnt5a, and Lrp6) to reduce the expressions of these genes. The inhibition of miR-12200-5p would almost fully alleviate postmenopausal osteoporosis. CONCLUSION: MiR-12200-5p could strongly repress osteoblast differentiation and bone formation by targeting multiple members of the Wnt signaling pathway simultaneously. The study supplemented the theoretical and experimental basis for researching the mechanism of osteogenic differentiation and inspired the development of novel therapeutic strategies for osteoporosis.

The combined role of PET/CT metabolic parameters and inflammatory markers in detecting extensive disease in small cell lung cancer.

The combined role of inflammatory markers [including neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), monocyte/lymphocyte ratio (MLR), and systemic immune-inflammation index (SII)] and PET/CT metabolic parameters [including maximum standardized uptake value (SUVmax), mean standardized uptake value (SUVmean), metabolic tumor volume (MTV), and TLG (total lesion glycolysis)] at baseline in evaluating the binary stage [extensive-stage disease (ED) and limited-stage disease (LD)] of small cell lung cancer (SCLC) is unclear. In this study, we verified that high metabolic parameters and inflammatory markers were related to the binary stage of SCLC patients, respectively (p < 0.05). High inflammatory markers were also associated with high MTV and TLG in patients with SCLC (p < 0.005). Moreover, the incidences of co-high metabolic parameters and inflammatory markers were higher in ED-SCLC (p < 0.05) than those in LD-SCLC. Univariate logistic regression analysis demonstrated that Co-high MTV/NLR, Co-high MTV/MLR, Co-high MTV/SII, Co-high TLG/NLR, Co-high TLG/MLR, and Co-high TLG/SII were significantly related to the binary stage of SCLC patients (p = 0.00). However, only Co-high MTV/MLR was identified as an independent predictor for ED-SCLC (odds ratio: 8.67, 95% confidence interval CI: 3.51-21.42, p = 0.000). Our results suggest that co-high metabolic parameters and inflammatory markers could be of help for predicting ED-SCLC at baseline. Together, these preliminary findings may provide new ideas for more accurate staging of SCLC.

Kaempferide enhances antioxidant capacity to promote osteogenesis through FoxO1/ß-catenin signaling pathway.

BACKGROUND: Forkhead box O1 (FoxO1)/ß-catenin signaling pathway is a main oxidative defense pathway, which plays essential roles in the regulation of osteoporosis (OP). The natural products possess quality therapeutic effects and few side effects. It is used as a novel strategy in the treatment of OP. However, there is no systematic study in the natural antioxidant drug based on the FoxO1/ß-catenin signaling pathway. This paper aims to discover pro-osteogenesis natural antioxidants for the prevention and treatment of OP. METHODS: Systems pharmacology; combined with reverse drug targeting, systems-ADME process, network analysis and molecular docking, was used to screen natural antioxidants based on the FoxO1/ß-catenin signaling pathway. Then in vitro experiments were performed to evaluate the osteogenesis effects of screened natural antioxidants. RESULTS: Kaempferide was screened as the most potential antioxidant to improve osteogenesis by the regulation of the FoxO1/ß-catenin signaling pathway. In vitro experiments showed that kaempferide significantly increased the expression of antioxidant genes and promoted osteogenic differentiation. Furthermore, kaempferide also improved the osteogenic differentiation inhibited by H2O2 through the enhancement of antioxidant capacity. Notably, kaempferide promoted cell antioxidant capacity by the increased nuclear translocation of FoxO1 and ß-catenin. CONCLUSIONS: These findings suggest that kaempferide is the natural antioxidant to promote osteogenesis effectively through the FoxO1/ß-catenin signaling pathway. Natural antioxidant therapy maybe a promising strategy for the prevention and treatment of OP.

Systems pharmacology dissection of action mechanisms for herbs in osteoporosis treatment.

Objective: Osteoporosis has become the biggest cause of non-fatal health issue. Currently, the limitations of traditional anti-osteoporosis drugs such as long-term ill-effects and drug resistance, have raised concerns toward complementary and alternative therapies, particularly herbal medicines and their natural active compounds. Thus, this study aimed to provide an integrative analysis of active chemicals, drug targets and interacting pathways of the herbs for osteoporosis treatment. Methods: Here, we introduced a systematic pharmacology model, combining the absorption, distribution, metabolism, and excretion (ADME) screening model, drug targeting and network pharmacology, to probe into the therapeutic mechanisms of herbs in osteoporosis. Results: We obtained 86 natural compounds with favorable pharmacokinetic profiles and their 58 targets from seven osteoporosis-related herbs. Network analysis revealed that they probably synergistically work through multiple mechanisms, such as suppressing inflammatory response, maintaining bone metabolism or improving organism immunity, to benefit patients with osteoporosis. Furthermore, experimental results showed that all the five compounds (calycosin, asperosaponin VI, hederagenin, betulinic acid and luteolin) enhanced osteoblast proliferation and differentiation in vitro, which corroborated the validity of this system pharmacology approach. Notably, gentisin and aureusidin among the identified compounds were first predicted to be associated with osteoporosis. Conclusion: Herbs and their natural compounds, being characterized as the classical combination therapies, might be engaged in multiple mechanisms to coordinately improve the osteoporosis symptoms. This work may contribute to offer novel strategies and clues for the therapy and drug discovery of osteoporosis and other complex diseases.

Evaluation of decreased uptake of 18F-fluorodeoxyglucose in the cerebral cortex of patients with intracranial non-Hodgkin's lymphoma lesions through PET/CT.

BACKGROUND: This study aims to investigate whether basal ganglia can be utilised as reference for measuring ratios between standardised uptake values (SUV), namely, SUVmax and SUVmean, of 18F-fluoro-deoxyglucose (18F-FDG) uptake, in the ipsilateral cerebral cortex and basal ganglia of patients with non-Hodgkin's lymphoma lesions. METHODS: Fifty-three patients with pathologically confirmed highly metabolic non-Hodgkin's lymphoma were retrospectively analysed; these patients were subjected to treatment strategy and underwent positron emission tomography/computed tomography (PET/CT). Nineteen of the patients who improved after the treatment were re-examined by PET/CT. PET/CT simultaneity was performed on 12 normal volunteers as the control group. SUVmax and SUVmean in the cerebral cortex and basal ganglia (head of caudate nucleus and lenticular nucleus) were evaluated by ganglia was also calculated. The ratios were compared between patients and the control group as well as before and after the treatment. RESULTS: The ratios between the SUVmax of 18F-FDG uptake were 1.03±0.16 (range 0.50-1.51, left) in the ipsilateral cerebral cortex and 1.02±0.16 (range 0.50-1.31, right) in the basal ganglia of 53 patients with non-Hodgkin's lymphoma; the corresponding ratios in the 12 controls were 1.09±0.11 (range 0.99-1.21, left) and 1.09±0.09 (range 1.00-1.24, right), respectively. The ratios between the SUVmean and 18F-FDG uptake in the ipsolateral cerebral cortex and basal ganglia were 0.76±0.09 (range 0.50-1.07, left) and 0.76±0.09 (range 0.48-0.98, right), respectively, which were lower than those in the control group [0.93±0.06 (range 0.83-0.99, left) and 0.92±0.05 (range 0.84-0.99, right), respectively]. For patients effectively treated, the ratios between the SUVmax in the ipsolateral cerebral cortex and basal ganglia were 1.08±0.13 (range 0.94-1.36, left) and 1.08±0.13 (range 0.88-1.31, right) before the treatment; these values were similar to 1.11±0.13 (range 0.85-1.36, left) and 1.09±0.11 (range 0.90-1.32, right) obtained in the ipsolateral cerebral cortex and basal ganglia of the patients after the treatment, respectively. The ratios between the SUVmean in the ipsolateral cerebral cortex and basal ganglia were 0.78±0.06 (range 0.68-0.93, left) and 0.78±0.06 (range 0.69-0.95, right) in the patients before the treatment; these ratios were lower than those in post-treatment patients [0.90±0.06 (range 0.74-1.00, left) and 0.90±0.07 (range 0.72-1.00, right), respectively], respectively. CONCLUSION: High levels of 18F-FDG metabolism in patients with non-Hodgkin's lymphoma may decrease glucose uptake in the cerebral cortex (diversion of 18F-FDG from the cerebral tissue to the lymphoma tissue); this phenomenon may be reversed with effective therapy for lymphoma. The ratio with 18F-FDG metabolism in basal ganglia could be used as reference to quantify and monitor glucose metabolism in cerebral tissues during the course of lymphoma.

[Impact of MRI-CT image registration on target delineation of postoperative radiotherapy for gliomas].

BACKGROUND & OBJECTIVE: Magnetic resonance imaging (MRI) can well distinguish soft tissue, but its usage in radiotherapy for brain tumors was limited by its image distortion and lack of electron density for dosage calculation. The registration of MRI with computed tomography (CT) could resolve this problem. This study was to investigate the registration accuracy of MRI-CT image, and compare the change of center position, clinical target volume (CTV) and volume of organs at risk (OARs). METHODS: Nine glioma patients were examined by MRI and CT after operation. The registration accuracy of MRI-CT was evaluated. The changes of CTV, volume of OARs, and center position were investigated by the method of volume and center of geometry (COG) respectively. COG was used to measure the distance between center positions on MRI-CT registration image and CT location image. The volume method was used to measure the percentage of the volume of registration area (VMRI-CT) to the total volume (VMRI+CT). RESULTS: The registration accuracy by the method of artificial landmarks was less than 1.5 mm, which reached the error requirement for brain tumors. No significant change of volume of OARs was found (P>0.05). Of the 9 patients, 8 had decreased CTV (by 13.85%-73.59%) on MRI-CT registration image and 1 had increased CTV (by 10.35%). The difference in mean CTV between the 2 groups was significant (P<0.05). The change of central position of CTV was the largest [(8.74+/-6.60) mm], those of 2 eyes were the next [(5.25+/-2.38) mm for the left eyeû (5.65+/-2.56) mm for the right eye], and that of the brain stem was the least [(1.83+/-1.06) mm]. CONCLUSIONS: By the method of artificial landmarks, MRI-CT has better registration accuracy. MRI-CT registration can reduce the uncertainty of CTV delineation in radiation treatment planning for glioma patients after operation.