Hedgehog signaling pathway affects the sensitivity of hepatoma cells to drug therapy through the ABCC1 transporter.

The poor response to drug therapy often seen in hepatocellular carcinoma requires insight into the molecular interplay responsible for intrinsic or acquired drug resistance. We previously demonstrated that the CD133-/EpCAM- subpopulation of the Huh-7 hepatoma cell line features aberrant activation of the hedgehog signaling (Hh) pathway and chemoresistance. The prevailing hypothesis of the present study is that hedgehog signaling may govern expression of ATP-binding cassette (ABC) transporters, which are responsible for drug resistance in the CD133-/EpCAM- subpopulation. Our aim is to reveal the molecular interplay in the mediation of drug resistance with a newly established Huh-7 subpopulation featuring high Hh signaling activity and drug resistance. In this study, chemoresistance was determined in a newly established Huh-7-DN subpopulation featuring the CD133-/EpCAM- surface marker profile, aberrant expression of Hh pathway, and epithelial-mesenchymal transition (EMT). Expression of ABC transporter proteins (ABCB1, ABCC1, and ABCG2) and Hh transcription factor Gli-1/2 was evaluated with and without Hh signaling antagonists LDE225 or itraconazole. We found that hedgehog signaling activity as determined by transfection with a Gli-Lux reporter cassette and gene expression levels tended to increase from Huh-7 CD133+/EpCAM+ to CD133-/EpCAM-, and the highest levels were found in Huh-7-DN cells. The Huh-7-DN subpopulation exhibited characteristics of EMT as evidenced by increased expression of vimentin and loss of E-cadherin. Sorafenib significantly inhibited the viability of all subpopulations except the Huh-7-DN subpopulation. Compared with other sorafenib-sensitive subpopulations, the Huh-7-DN subpopulation showed enhanced expression of Hh transcription factor Gli-2 and ABCC1 transporter protein. Silencing Gli-2 by lentivirus harboring shRNA against Gli-2 or LDE225 significantly suppressed expression of Gli-2 and ABCC1 genes in Huh-7-DN subpopulation. In conclusion, aberrant hedgehog signaling activation is linked to poor differentiation, epithelial-mesenchymal transition, and chemoresistance in the Huh-7-DN subpopulation. Hedgehog signaling transcription factor Gli-2 appears to be the primary regulator for drug sensitivity of hepatoma through the ABCC1 transporter.

Influences of Heider Balance on Knowledge, Attitude, Practice, and Quality of Life in Bladder Cancer Patients after Urinary Diversion.

Objective: To explore the influences of Heider balance on knowledge, attitude, practice (KAP), and quality of life in bladder cancer patients after urinary diversion. Methods: A set of bladder cancer patients after urinary diversion in our hospital from January 2016 to December 2020 were included in this study. Patients who received out-hospital intervention based on Heider balance were included in the observation group (85 cases). Meanwhile, patients who received routine out-hospital intervention were included in the control group (85 cases), and these patients matched with the observation group by gender, age, and education level. The scores of KAP, WHO quality of life-100 (WHOQOL-100) before discharge and at 6 months after discharge, and the rate of complications were compared in the two groups. Results: At 6 months after discharge, the score of these items of KAP including basic knowledge of disease, procedure of pouch replacement, dealing with pouch leakage, skin care of stoma, purchase and storage of pouch, dealing with stoma complications, optimistic mentality for disease, optimistic mentality for stoma, trust in medical staff, willingness to correct bad habits, confidence in maintaining health behavior, maintaining in health dietary habit, maintaining in health behavior, learning from relevant books, learning from relevant videos, experienced in pouch replacement, and experienced in care of stoma of the observation group were significantly higher than those of the control group (t = 6.144, 9.366, 3.129, 3.809, 4.173, 5.923, 2.788, 8.871, 3.291, 10.797, 7.067, 7.805, 3.828, 9.454, 2.827, 4.059, and 8.662, respectively, all P < 0.05). The scores of 16 items of WHOQOL-100 such as energy and fatigue, sleep and rest, positive feelings, thinking, learning, memory and concentration, self-esteem, body image and appearance, negative feelings, mobility, activities of daily living, dependence on medical support, personal relationships, social support, health and social care: availability and quality, opportunities to get new information/skills, opportunities for recreation and leisure, and quality of life from viewpoint in the observation group were significantly higher than those in the control group (t = 2.666, 2.571, 2.961, 3.453, 4.279, 2.781, 3.775, 4.807, 5.850, 4.194, 3.324, 3.873, 5.118, 3.244, 2.956, and 4.218, respectively, all P < 0.05). The rate of complications of the observation group was significantly lower than that of the control group (x 2 = 5.829, P < 0.05). Conclusion: The Heider balance can help to reduce the rate of complications, improve knowledge, attitude, practice, and quality of life in urinary diversion patients. These merits make it an attractive approach in guidance of out-hospital intervention.

Aberrant miR-339-5p/neuronatin signaling causes prodromal neuronal calcium dyshomeostasis in mutant presenilin mice.

Mushroom spine loss and calcium dyshomeostasis are early hallmark events of age-related neurodegeneration, such as Alzheimer's disease (AD), that are connected with neuronal hyperactivity in early pathology of cognitive brain areas. However, it remains elusive how these key events are triggered at the molecular level for the neuronal abnormality that occurs at the initial stage of disease. Here, we identify downregulated miR-339-5p and its upregulated target protein, neuronatin (Nnat), in cortex neurons from the presenilin-1 M146V knockin (PSEN1-M146V KI) mouse model of familial AD (FAD). Inhibition of miR-339-5p or overexpression of Nnat recapitulates spine loss and endoplasmic reticulum calcium overload in cortical neurons with the PSEN1 mutation. Conversely, either overexpression of miR-339-5p or knockdown of Nnat restores spine morphogenesis and calcium homeostasis. We used fiber photometry recording during the object-cognitive process to further demonstrate that the PSEN1 mutant causes defective habituation in neuronal reaction in the retrosplenial cortex and that this can be rescued by restoring the miR-339-5p/Nnat pathway. Our findings thus reveal crucial roles of the miR-339-5p/Nnat pathway in FAD that may serve as potential diagnostic and therapeutic targets for early pathogenesis.

MiR-93 functions as a tumor promoter in prostate cancer by targeting disabled homolog 2 (DAB2) and an antitumor polysaccharide from green tea (Camellia sinensis) on their expression.

Our previous work has demonstrated that the role of miR-93 in prostate cancer (PC) progression. The aim of this study was to determine the downstream gene regulated by miR-93 and the molecular mechanisms underlying its roles in PC. Bioinformatics analysis and luciferase reporter assays predicted disabled homolog 2 (DAB2) as a direct target gene of miR-93. Real time quantitative polymerase chain reaction (qRT-PCR) and Western blot analysis revealed that DAB2 was tumor repressor in PC cells, and its mRNA expression was negatively correlated with miR-93 in PC tissues. Gain and loss of function experiments also indicated DAB2 overexpression significantly suppressed PC cells proliferation, invasion and migration, while knockdown of its expression came to the opposite effect. Furthermore, a rescue experiment indicated miR-93 directly regulated PC cell growth and migration, as well as AKT and ERK activation by targeting DAB2. Additionally, antitumor effect of a Green tea polysaccharide (GTP) on PC-3 cells could be achieved by increasing DAB2 protein expression and inactivating AKT and ERK1/2 signaling. Our study suggests that miR-93 promoted PC progression and metastasis by repressing DAB2 to activate Akt/ERK1/2 pathway, and elevation of DAB2 and inactivation of Akt/ERK1/2 might be a potential therapeutic target for PC by GTP.

Anti-tumor activity and the mechanism of a green tea (Camellia sinensis) polysaccharide on prostate cancer.

In this study, a homogeneous polysaccharide (GTP), with a molecular weight of 7.0 × 104 Da, was isolated from Green tea, which was only composed of glucose. The antitumor effects of GTP on prostate cancer (PC) cell line along with the possible mechanism was examined. First, we investigate the potential role of microRNA-93 (miR-93) in PC progression. Our results showed that miR-93 was significantly upregulated in human PC tissues and several PC cell lines, and its overexpression was correlated with poor survival in PC patients. Furthermore, functional analysis showed that miR-93 overexpression promoted the migration, invasion and proliferation of PC-3 cells transfected with miR-93 mimics, while its knockdown displayed an opposite result in DU145 cells following miR-93 inhibitor transfection. Additionally, in vivo tumorigenic studies on nude mice confirmed that miR-93 mimic treatment accelerated the growth of PC-3 xenograft tumors. As expected, GTP (25, 50 and 100 µg/ml) inhibited growth of PC-3 cells via inducing apoptosis, which was achieved by elevation of bax/bcl-2 ratio and caspae-3 protein expression, as well as a decrease of miR-93. Thus, miR-93 may be a potential therapeutic target by GTP for PC therapy.