Stress, Vascular Smooth Muscle Cell Phenotype and Atherosclerosis: Novel Insight into Smooth Muscle Cell Phenotypic Transition in Atherosclerosis.

PURPOSE OF REVIEW: Our work is to establish more distinct association between specific stress and vascular smooth muscle cells (VSMCs) phenotypes to alleviate atherosclerotic plaque burden and delay atherosclerosis (AS) progression. RECENT FINDING: In recent years, VSMCs phenotypic transition has received significant interests. Different stresses were found to be associated with VSMCs phenotypic transition. However, the explicit correlation between VSMCs phenotype and specific stress has not been elucidated clearly yet. We discover that VSMCs phenotypic transition, which is widely involved in the progression of AS, is associated with specific stress. We discuss approaches targeting stresses to intervene VSMCs phenotypic transition, which may contribute to develop innovative therapies for AS.

PLAC8 promotes adriamycin resistance via blocking autophagy in breast cancer.

Adriamycin (ADM) is currently one of the most effective chemotherapeutic agents in breast cancer treatment. However, growing resistance to ADM could lead to treatment failure and poor outcome. PLAC8 was reported as a novel highly conserved protein and functioned as an oncogene or tumour suppressor in various tumours. Here, we found higher PLAC8 expression was correlated with worse outcome and aggressive phenotype in breast cancer. Breast cancer patients with higher PLAC8 expression showed potential ADM resistance. In vitro experiments further confirmed that PLAC8 inhibited by siRNA or enforced overexpression by infecting pcDNA3.1(C)-PLAC8 plasmid correspondingly decreased or increased ADM resistance. Subsequently, we demonstrated that ectopic PLAC8 expression in MCF-7/ADMR cell blocked the accumulation of the autophagy-associated protein LC3 and resulted in cellular accumulation of p62. Rapamycin-triggered autophagy significantly increased cell response to ADM, while the autophagy inhibitor 3-MA enhanced ADM resistance. 3-MA and PLAC8 could synergistically cause ADM resistance via blocking the autophagy process. Additionally, the down-regulation of p62 by siRNA attenuated the activation of autophagy and PLAC8 expression in breast cancer cells. Thus, our findings suggest that PLAC8, through the participation of p62, inhibits autophagy and consequently results in ADM resistance in breast cancer. PLAC8/p62 pathway may act as novel therapeutic targets in breast cancer treatment and has potential clinical application in overcoming ADM resistance.

Upregulation of lincRNA-p21 in thoracic aortic aneurysms is involved in the regulation of proliferation and apoptosis of vascular smooth muscle cells by activating TGF-ß1 signaling pathway.

OBJECTIVE: Long intergenic noncoding RNA-p21 (lincRNA-p21) has been proved in the pathogenesis of aortic aneurysms, while its functionality in thoracic aortic aneurysms (TAA) and the mechanism of function remains unclear. Therefore our study aimed to investigate the role of lincRNA-p21 in TAA. METHODS: Aortic media specimens and blood samples were collected from both patients with TAA and healthy controls. Expression of lincRNA-p21 in those tissues was detected by reverse-transcription quantitative polymerase chain reaction (qRT-PCR). Diagnostic values of lincRNA-p21 in aortic media and blood for TAA were evaluated by receiver operating characteristic curve analysis. LincRNA-p21 overexpression human vascular smooth muscle cells (VSMCs) were prepared and the effects of lincRNA-p21 overexpression on cell proliferation and apoptosis were explored by cell counting kit-8 assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, respectively. Expression of lincRNA-p21 and transforming growth factor ß1 (TGF-ß1) in VSMCs with different treatment was detected by qRT-PCR and Western blot analysis, respectively. RESULTS: Expression of lincRNA-p21 in aortic media tissues and blood was significantly upregulated in TAA patients than in healthy controls. Expression of lincRNA-p21 in aortic media and blood can be used to effectively distinguish TAA patients form healthy controls. LincRNA-p21 overexpression inhibited proliferation and promoted apoptosis of VSMCs, while TGF-ß1 inhibitor reduced those effects. LincRNA-p21 overexpression upregulated TGF-ß1 expression, while TGF-ß1 activator showed no significant effects on lincRNA-p21 expression in VSMC. CONCLUSION: LincRNA-p21 participates in TAA by regulating the proliferation and apoptosis of VSMCs through the activation of TGF-ß1 signaling pathway.

The Expression Profiles of MicroRNA Let-7a in Peripheral Blood Mononuclear Cells from Patients of Gastric Cancer with Neoadjuvant Chemotherapy.

BACKGROUND: The microRNA let-7a contains three copies of genes and is associated with various types of cancer, including gastric cancer. In this study, we aim to investigate the differential expression patterns of microRNA let7a in PBMCs from patients of gastric cancer (GC) before and after neoadjuvant chemotherapy. METHODS: The differential expression levels of let-7a were detected in PBMCs from 22 patients with GC before and after neoadjuvant chemotherapy by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The statistical analysis was performed with t-test and one-way AVOVA. RESULTS: MicroRNA let-7a level was significantly decreased in patients with GC after neoadjuvant chemotherapy (p < 0.05). In patients with GC, microRNA let-7a has different expression patterns with different neoadjuvant chemotherapy cycles. CONCLUSIONS: Our study demonstrated that microRNA let-7a was expressed differentially in patients with GC before and after neoadjuvant chemotherapy. These data supported that microRNA let-7a may have a potential role in efficacy assessment in patients with GC with neoadjuvant chemotherapy.

Structure and function of corneal surface of mudskipper fishes.

Vertebrate corneal epithelium cell plays an important role for imaging, and the cell density, together with the appearance or type of affiliated microstructures, is considered as a result of evolution adapting to alternate terrestrial or aquatic environment. In this paper, we investigated the corneal cells of both larvae and adult amphibious mudskippers Boleophthalmus pectinirostris and Periophthalmus magnuspinnatus, to testify the relationship between morphology and function. The cell density values of the two species were 31,137 and 31,974 cells per mm(2) in larvae and then significantly decreased to 15,826 and 25,954 cells per mm(2) in adult (p < 0.001), respectively, which could be explained as the habitat change from aquatic to different degrees of terrestrial environment. The corneal epithelium cells were ridge type in larvae and differentiated into ridge type and reticular type in adult P. magnuspinnatus and ridge type, reticular type and ridge-reticular type in adult B. pectinirostris. Four kinds of microstructures as microridge, microvilli, microplicae and microhole appeared in both species. The difference of microridge width and its separation indicated that a dense cell connection was requested in a saltier and more terrestrial environment.

Adaptive evolution of antioxidase-related genes in hypoxia-tolerant mammals.

To cope with the damage from oxidative stress caused by hypoxia, mammals have evolved a series of physiological and biochemical traits, including antioxidant ability. Although numerous research studies about the mechanisms of hypoxia evolution have been reported, the molecular mechanisms of antioxidase-related genes in mammals living in different environments are yet to be completely understood. In this study, we constructed a dataset comprising 7 antioxidase-related genes (CAT, SOD1, SOD2, SOD3, GPX1, GPX2, and GPX3) from 43 mammalian species to implement evolutionary analysis. The results showed that six genes (CAT, SOD1, SOD2, SOD3, GPX1, and GPX3) have undergone divergent evolution based on the free-ratio (M1) model. Furthermore, multi-ratio model analyses uncovered the divergent evolution between hypoxic and non-hypoxic lineages, as well as various hypoxic lineages. In addition, the branch-site model identified 9 positively selected branches in 6 genes (CAT, SOD1, SOD2, SOD3, GPX2, and GPX3) that contained 35 positively selected sites, among which 31 positively selected sites were identified in hypoxia-tolerant branches, accounting for 89% of the total number of positively selected sites. Interestingly, 65 parallel/convergent sites were identified in the 7 genes. In summary, antioxidase-related genes are subjected to different selective pressures among hypoxia-tolerant species living in different habitats. This study provides a valuable insight into the molecular evolution of antioxidase-related genes in hypoxia evolution in mammals.

Evaluation of the upper lakes of the Erhai watershed in China based on water quality, land cover, and macrobenthic invertebrates.

In this study, three lakes, Cibi Hu, Haixi Hai, and Xi Hu, in the upper reaches of the main inflow rivers in the northern part of Erhai Lake were selected as the research objects. Based on the water environment monitoring indicators, land cover data, and lake macrobenthic community observation data, the non-parametric Kruskal-Wallis test, spatial analysis and community structure analysis were used to quantitatively assess the water environment and ecological status of the lakes. Using the Pollution Tolerance Index (PTI), the potential utility of macroinvertebrate communities as indicators of water ecological quality was investigated. The results showed that Cibi Hu and Haixi Hai have similar characteristics on water environmental quality. The physical and chemical indexes of water quality, the land cover of the lake catchment area, and the PTI index of the benthic community showed that Xi Hu was the most affected by human disturbance; the water ecological condition was the worst; and the environmental protection pressure was the greatest. In general, PTI analysis based on benthic fauna is convenient and can reflect the basic conditions of the aquatic benthic environment keenly, which is worthy of promotion.

Exposure status, spatial variation, and health risk assessment of selected heavy metal(loid)s in common commercial fish species of the Beibu Gulf.

Fishes are considered as biological indicators of heavy metal(loid)s pollution. In this study, contents of seven heavy metal(loid)s, including Cu, Cr, Cd, Pb, Zn, As, and Hg, in the muscles of ten common fish species in the Beibu Gulf were analyzed to figure out the pollutants status and their health risk. Results showed all species were largely contaminated by arsenic. Under conservative estimation scenario, target hazard quotient and health index revealed no health risk of species except Alepes kleinii. Under pessimistic estimation scenario, target cancer risk and estimated daily intake showed that, except Saurida undosquamis, Saurida tumbil, and Trachinotus ovatus, the remaining species were at risk of causing cancer for their consumers. Daily intake of arsenic and mercury in most species by residents in the Beibu Gulf exceeded provisional maximum tolerable amount recommended by FAO, suggesting the need of moderate consumption of these species.

Challenging Isodimorphism Concepts: Formation of Three Crystalline Phases in Poly(hexamethylene-ran-octamethylene carbonate) Copolymers.

In this work, poly(hexamethylene-ran-octamethylene carbonate) copolycarbonates were synthesized by melt polycondensation in a wide range of compositions. The copolymers displayed some of the characteristic isodimorphic thermal behavior, such as crystallization for all the compositions and a pseudoeutectic behavior of the melting temperature (Tm) versus composition. The pseudoeutectic point was located at 33 mol % poly(octamethylene carbonate) (POC) content (i.e., corresponding to the PH67O33C copolymer). Surprisingly, the crystallinities (Xc) for a wide range of copolymer compositions were higher than those of the parent components, a phenomenon that has not been observed before in isodimorphic random copolymers. The structural characterization, performed by wide-angle X-ray scattering (WAXS) and small-angle X-ray scattering experiments, revealed unexpected results depending on composition. On the one hand, the poly(hexamethylene carbonate) (PHC)- and POC-rich copolymers crystallize in PHC- and POC-type crystals, as expected. Moreover, upon cooling and heating, in situ WAXS experiments evidenced that these materials undergo reversible solid-solid transitions [δ-α (PHC) and δ-α-ß (POC)] present in the parent components but at lower temperatures. On the other hand, a novel behavior was found for copolymers with 33-73 mol % POC (including the pseudoeutectic point), which are those with higher crystallinities than the parent components. For these copolymers, a new crystalline phase that is different from that of both homopolymers was observed. The in situ WAXS results for these copolymers confirmed that this novel phase is stable upon cooling and heating and does not show any crystallographic feature of the parent components or their solid-solid transitions. FTIR experiments confirmed this behavior, revealing that the new phase adopts a polyethylene-like chain conformation that differs from the trans-dominant ones exhibited by the parent components. This finding challenges the established concepts of isodimorphism and questions whether a combination of crystallization modes (isodimorphism and isomorphism) is possible in the same family of random copolymers just by changing the composition.

[Prognostic effect of postoperative resection-margin status for intraoperative positive resection margins in patients with advanced gastric cancer].

OBJECTIVES: To investigate prognostic effect of postoperative resection-margin status for intraoperatively positive resection margin in advanced gastric cancer and discuss the treatment choice for intraoperatively positive resection margins. METHODS: A retrospective study was investigated in 64 advanced gastric cancer patients with positive resection margin after potentially curative resection. The survival between 50 patients who was re-excised to a negative resection margin (NR group) and 14 patients who were left with positive resection margin (PR group) was compared. Prognostic factors were analyzed using univariate and multivariate Cox regression model analysis. RESULTS: The median survival in the PR group was 17.0 months (95%CI: 11.6 - 22.4) as compared with 23.0 months (95%CI: 20.5 - 25.5) in the NR group (P = 0.045). However, resection-margin status lost significance on multivariate analysis. In the subgroup of D2 lymphadenectomy, the median survival in the PR group and NR group were 17.0 months (95%CI: 12.0 - 22.0) and 24.0 months (95%CI: 19.8 - 28.1) respectively; multivariate analysis further identified resection margin status as an independent prognostic factor. CONCLUSIONS: Re-excision for intraoperatively positive margin to negative margin improves the prognosis of the patients with advanced gastric cancer, and re-excision is the first choice when intraoperative frozen section detects a positive margin. Routine frozen section of resection margin should be mandatory in all advanced gastric cancer undergoing potentially curative surgery.

Prognostic Significance of Neutrophil-to-Lymphocyte Ratio and C-Reactive Protein/Albumin Ratio in Luminal Breast Cancers With HER2-Negativity.

Purpose: This study was determined to evaluate the prognostic value of neutrophil-to-lymphocyte ratio (NLR) and C-reactive protein/albumin ratio (CAR) prior to surgery in luminal breast cancers (BC) with HER2-negativity. Methods: The clinical data of 708 HER2-negative luminal BC patients from January 2013 to December 2016 were retrospectively collected and analyzed. The optimal cut-off value of NLR and CAR were determined via receiver operating characteristic (ROC) curve. The disease-free survival (DFS) and cancer specific survival (CSS) rates were estimated using the Kaplan-Meier method. Cox univariate and multivariate proportional hazards regression models were performed to identify significant predictors of DFS and CSS simultaneously. Results: The mean age of the patients diagnosed was 52.43 years (range, 15-95 years), and the median follow-up was 62.71 months (range, 12-92 months). Univariate and multivariate analysis confirmed that NLR ≥2.2 was significantly associated with worse DFS (HR=2.886, 95%CI=1.756-4.745, p<0.001), and same results were obtained in terms of CSS (HR=3.999, 95%CI=2.002-7.987, p<0.001). Similarly, CAR ≥0.07 was independently and significantly associated with poor DFS (HR=3.858, 95%CI=2.346-6.345, p<0.001) and CSS (HR=6.563, 95%CI=3.558-12.106, p<0.001). Conclusion: Preoperative evaluation of NLR and CAR were significant and independent prognostic indicators for luminal breast cancers with HER2-negativity.

The influence of preoperative biopsy on the surgical method in breast cancer patients: a single-center experience of 3,966 cases in China.

BACKGROUND: The National Surgical Adjuvant Breast and Bowel Project (NSABP) B32 trial reported that the detection rate of sentinel lymph nodes by core needle biopsy (CNB) is higher than that by segmental resection. However, there are few reports regarding the detection rate of sentinel lymph nodes by vacuum-assisted breast biopsy (VABB). Therefore, we analyzed the impact of preoperative biopsy methods on the surgical modes of 3,966 patients with breast cancer in our center. METHODS: In total, 3,966 female breast cancer patients [clinical tumor node metastasis (TNM) stage I-III] were enrolled in this study. Preoperative pathological diagnosis methods included fine needle aspiration (FNA) biopsy, CNB, excision biopsy, and VABB. According to the time of diagnosis. The data were analysis by chi square test, variance analysis and the Kaplan-Meier time series in SPSS 22.0. RESULTS: There was a decrease in the number of patients that underwent excision biopsy (7.3% to 2.7%) and intraoperative freezing (89.4% to 28.9%) over time, while CNB exhibited an increasing trend (1.6% to 55.3%). The positive rates of VABB, CNB, excision biopsy, and FNA were 99.5%, 97.1%, 97.9%, and 82.2%, respectively, and the false negative rates were 0%, 1.8%, 0.34%, and 8.9%, respectively. The overall breast-conserving rate was 36.7%, while the breast-conserving rate for VABB was 57.1%. The axillary sentinel lymph node biopsy rate of cN0 patients was 48.3%, and the intraoperative frozen group (36.7%) and excision biopsy group (39.5%) were lower than the CNB (57.1%) and VABB (77.9%) groups. Until December 2019, there were 350 cases with tumor recurrence or metastasis. The methods of biopsy were not correlated to the cumulative survival time. CONCLUSIONS: Changes to the diagnosis and treatment of breast cancer has a profound impact on the method of tumor biopsy. VABB biopsy offers advantages such as accurate diagnosis, a greater volume of tissue taken at one time, minimally invasive and repeatable, and does not affect the surgical approach and prognosis of patients. It will gradually become the primary method of preoperative pathological evaluation of breast cancer.

Pegylated Liposomal Doxorubicin Versus Epirubicin as Adjuvant Therapy for Stage I-III Breast Cancer.

Background: Conventional anthracyclines, like epirubicin, are cornerstone drugs for breast cancer treatment of all stages, but their cumulative toxicity could cause life-threatening side effects. Pegylated liposomal doxorubicin (PLD), an effective anti-breast cancer drug, has lower toxicity than conventional anthracyclines. This retrospective study compared the efficacy and toxicity profiles between PLD and epirubicin as adjuvant therapy for breast cancer. Patients and Methods: A total of 1,471 patients diagnosed with stage I-III breast cancer between 2000 and 2018 were included in this study, among which 661 were treated with PLD and 810 with epirubicin, with 45.9 months as the median follow-up time. Anti-breast cancer efficacy was assessed with overall survival (OS) and disease-free survival (DFS), while cardiac toxicity was assessed with left ventricular ejection fraction (LVEF) and electrocardiogram (ECG). Results: The Kaplan-Meier method and Cox proportional hazards model revealed that there was no statistical difference in OS or DFS between patients treated with PLD and epirubicin, regardless of cancer stages or molecular subtypes (all p-values > 0.05). In addition, patients had significantly better LEVF and ECG data after adjuvant therapy with PLD (both p-values < 0.05). Conclusion: Based on the large sample size and the long follow-up time of this study, we conclude that PLD has a similar anti-breast cancer efficacy as epirubicin while inducing lower level of cardiac toxicity in Han Chinese. This study suggests that PLD-based adjuvant chemotherapy could be a better option than epirubicin for breast cancer patients especially with existing cardiac disease.

Evaluation of the diagnostic value of circulating tumor cells with CytoSorter® CTC capture system in patients with breast cancer.

PURPOSE: In this study, we aimed to investigate the viability of utilizing CytoSorter® system to detect circulating tumor cells (CTCs) and to evaluate the diagnostic value of CTCs in breast cancer (BC). METHODS: A total of 366 females patients suspected of having BC and 30 healthy female volunteers were enrolled in this study. CTCs were enriched by CytoSorter® , a microfluidic-based CTCs capturing platform. CTC detection was performed before operation or biopsy. Based on the biopsy results, patients were divided into two groups, namely patients with BC and patients with benign breast diseases (BBD). Patients with BBD and healthy volunteers were serving as controls. The correlation between CTC enumeration and patients' clinicopathological characteristics was evaluated. The receiver operating characteristic (ROC) curve was plotted to assess the diagnostic potency of CytoSorter® system in BC. RESULTS: Based on the biopsy results, 130 BC patients at different cancer stages and 236 patients with BBD were enrolled in the study. Seven subjects were dropped out from the study. CTCs were detected in 109 of 128 BC patients, in one of 29 healthy volunteers, and in 37 of 232 patients with BBD. Maximum CTC counts detected in BC patients, healthy volunteers, and patients with BBD were 8, 1, and 4, respectively. Statistical analysis showed CTCs could be used to distinguish BC patients from healthy volunteers and patients with BBD (P < .0001). Circulating tumor cells were statistically associated with patients' cancer stage (P = .0126), tumor size (tumor node metastasis [TNM] T stage, P = .0253), cancer type (invasive vs noninvasive, P = .0141), and lymph node metastasis (P = .0436). More CTCs were found in patients at advanced cancer stage or TNM T stage and in patients with invasive tumor or lymph node metastasis. Furthermore, CTC detection rates in BC patients at Tis and T1-4 stages were 50%, 81.67%, 91.07%, 100%, and 100%, respectively. When the CTC cut-off value was set to 2, the ROC curve gave an area under the curve (AUC) of 0.86 with a specificity and sensitivity of 95.4% and 76.56%, respectively. Taken together, CTCs could be used as a diagnostic aid in assistance of cancer screening and staging. CONCLUSION: Circulating tumor cells were successfully isolated in BC patients using CytoSorter® system. CTCs can be used to differentiate BC patients from the patients with BBD or healthy volunteers, and as a diagnostic aid for early cancer diagnosis and cancer staging.

Double-stranded RNA-dependent protein kinase-dependent apoptosis induction by a novel small compound.

The interferon-induced, double-stranded RNA-dependent protein kinase (PKR) can play critical roles in inhibiting virus replication and inducing apoptosis. To develop new agents that may inhibit viral replication or induce apoptosis in cancer cells via the PKR signaling pathway, we screened a chemical library for compounds that have differential cytotoxic effects on wild-type [mouse embryonic fibroblast (MEF)/PKR(+/+)] and PKR-knockout [MEF/PKR(-/-)] mouse embryonic fibroblast cells. We identified a synthetic compound, BEPP [1H-benzimidazole1-ethanol,2,3-dihydro-2-imino-a-(phenoxymethyl)-3-(phenylmethyl)-,monohydrochloride], that induces a cytotoxic effect more effectively in MEF/PKR(+/+) cells than in MEF/PKR(-/-) cells. BEPP also relatively effectively inhibited the growth of a human lung cancer cell line overexpressing PKR, compared with other cancer cell lines. In sensitive cells, BEPP induced apoptosis with activation of caspase-3. Treatment with BEPP led to increased phosphorylation of PKR and eIF2alpha, increased expression of BAX, and decreased expression of Bcl-2. BEPP-induced apoptosis was PKR dependent and was blocked by the adenovector expressing the dominant-negative PKR. Furthermore, pretreatment of HeLa cells at a noncytotoxic dose of BEPP effectively inhibited Vaccinia virus replication. Together, our results suggest that BEPP and its analogs may induce PKR-dependent apoptosis and inhibition of viral replication and that they can be a potential anticancer or anti-virus agent.

Adenoviral endoplasmic reticulum-targeted mda-7/interleukin-24 vector enhances human cancer cell killing.

We developed several adenoviral vectors designed to target MDA-7 expression to different subcellular compartments [endoplasmic reticulum (ER), mitochondria, nucleus, and cytosol] and evaluated their ability to enhance apoptosis. Adenoviral ER-targeted mda-7/interleukin-24 vector (Ad-ER-mda7) selectively and effectively inhibited the growth and proliferation of lung (A549 and H1299) and esophageal (Seg1 and Bic1) cancer cells by enhancing cell killing. Both Ad-mda7 and Ad-ER-mda7 activated a novel pathway of ER stress-induced apoptosis characterized by unregulated expression of phosphorylated JNK, phosphorylated c-Jun, and phosphorylated RNA-dependent protein kinase. Caspase-4 activation mediated Ad-mda7- and Ad-ER-mda7-induced cell death. In addition, Ad-mda7- and Ad-ER-mda7-mediated growth inhibition correlated with activation of ER molecular markers RNA-dependent protein kinase and JNK both in vitro (in Ad-mda7- or Ad-ER-mda7-treated lung cancer cells) and in vivo. These findings suggest that vectors targeting the ER (Ad-ER-mda7) may be more effective in cancer gene therapy possibly through more effective induction or ER stress pathways.

Therapeutic effect of the epidermal growth factor on diabetic foot ulcer and the underlying mechanisms.

The present study aimed to explore the therapeutic effect and underlying mechanism of epidermal growth factor (EGF) on the wound healing of diabetic foot ulcers (DFU). A total of 48 rabbits with DFU were randomly divided into 2 groups, comprising the treatment and control groups. Full-thickness skin (10×10 mm) was excised from the thigh of each rabbit. The wounds in the treatment group were treated with 100 mg/l EGF once a day for 1 month. The control group received no treatment. At 20 days following treatment, new granulation tissues that formed beyond the edge of the wound were collected for subsequent analysis. Tissues from rabbits in the treatment group produced a greater number of fibroblasts, which exhibited a fibroblastic morphology when compared with those in the control group. In the treatment group, a larger number of these fibroblasts were observed as clusters, and there were numerous blood vessels when compared with the control group. The fibroblasts in the control group exhibited an irregular morphology, contained fewer organelles and the surrounding collagenous fibers were sparse. These fibroblasts also demonstrated a disordered arrangement and it was revealed that the wound healed at a slower rate compared with the treatment group. Endogenous EGF mRNA detection revealed that there was a significant difference (P<0.05) in the relative gray value of EGF mRNA between the treatment (103.27±4.27) and control (63.88±4.36) groups. In conclusion, EGF may accelerate the healing of DFU, and exogenous EGF treatment may upregulate the expression of EGF mRNA in newly generated tissues.

The novel KLF4/PLAC8 signaling pathway regulates lung cancer growth.

Accumulating evidence suggests that placenta-specific 8 (PLAC8) plays an important role in normal cellular process and human diseases, including multiple types of human tumors, and its role is highly relied upon in cellular and physiologic contexts. However, there are no reports on its expression profile and biological roles during lung cancer development. In the current study, both the clinical implications and biological effects of PLAC8 in lung cancer (LC) progression were investigated, and we identified and described the novel Krüppel-like factor 4 (KLF4)/PLAC8 regulatory pathway in cancer progression. Elevated PLAC8 levels were positively correlated with tumor size, histological grade, and tumor node metasis (TNM) stage, and LC patients with high PLAC8 expression suffered poor outcomes. In vitro and in vivo assays further revealed that endogenous PLAC8 promoted cell proliferation and tumor formation. We also found downregulated PLAC8 protein in several LC cell lines following the induction of KLF4, and immunohistochemistry analysis of LC tissues by microarray indicated a potential inverse correlation between PLAC8 and KLF4 expression. Luciferase reporter analysis and chromatin immunoprecipitation assays determined that KLF4 negatively regulated PLAC8 promoter activity via directly binding to the promoter region. Furthermore, the growth inhibition resulting from KLF4 overexpression was partially rescued by ectopic PLAC8 expression. Together, our data uncovered a previously unidentified role of PLAC8 as a central mediator in LC progression. PLAC8 was transcriptionally repressed by KLF4, and the novel KLF4/PLAC8 axis may act as a promising candidate target for LC diagnosis and therapy.

MLN4924 neddylation inhibitor promotes cell death in paclitaxel-resistant human lung adenocarcinoma cells.

Acquired resistance to first-line chemotherapeutics, including paclitaxel (PTX), is a primary factor contributing to chemotherapy failure in non-small cell lung cancer (NSCLC) patients. Previous studies have identified that targeting NEDD8-activating enzyme (NAE) with MLN4924 effectively overcomes platinum resistance in preclinical models of ovarian cancer. However, the underlying mechanisms are yet to be fully elucidated. The present study demonstrates that the inhibition of the neddylation pathway with MLN4924 an NAE inhibitor inhibited protein neddylation, inactivated cullin-RING E3 ligase and exhibited a potent antiproliferative effect on PTX-resistant A549 and H460 cells (A549/PTX and H460/PTX). The application of MLN4924 promotes apoptosis and DNA damage in A549/PTX and H460/PTX cells. Additionally, MLN4924 abrogated the 3-dimensional growth potential of these cells and inhibited the formation of the A549/PTX and H460/PTX spheroids. Notably, combining MLN4924 with PTX did not exhibit synergy in PTX-resistant NSCLC cells. Taken together, the results of the current study suggest that MLN4924 may be utilized as an effective strategy for the treatment of PTX-resistant NSCLC.

KLF4 overcomes tamoxifen resistance by suppressing MAPK signaling pathway and predicts good prognosis in breast cancer.

Tamoxifen resistance represents a daunting challenge to the successful treatment for breast cancer. Krüppel-like factor 4 has critical roles in the development and progression of breast cancer, but its expression, function and regulation in the efficacy of TAM therapy in breast cancer have yet to be investigated. Here, we examined the clinical significance and biologic effects of KLF4 in breast cancer. Firstly, higher expression of KLF4 correlated with increased TAM sensitivity in breast cancer cells, and analysis of GEO datasets indicated that KLF4 expression was positively correlated with ERα and enhanced expression of KLF4 sensitized breast cancer patients to endocrine therapy. Knockdown of KLF4 in MCF-7 and BCAP37 cells led to increased TAM resistance, while ectopic KLF4 expression promoted the responsiveness to TAM in T47D and TAM-resistant MCF-7/TAM cells. Secondly, ectopic KLF4 overexpression suppressed MCF-7/TAM cell growth, invasion and migration. Moreover, KLF4 expression was down-regulated in breast cancer tumor tissues and high expression of KLF4 was associated with favorable outcomes. Mechanistically, KLF4 may enhance the responsiveness of breast cancer cells to TAM through suppressing mitogen-activated protein kinase (MAPK) signaling pathway. We found that ERK and p38 were more activated in MCF-7/TAM compared with MCF-7, and treatment with MAPK-specific inhibitors significantly suppressed cell viability. Knockdown of KLF4 activated ERK and p38 and drove MCF-7 cells to become resistant to TAM. Conversely, overexpression of KLF4 in MCF-7/TAM cells suppressed ERK and p38 signaling and resulted in increased sensitivity to TAM. Therefore, our findings suggested that KLF4 contributed to TAM sensitivity in breast cancer via phosphorylation modification of ERK and p38 signaling. Collectively, this study highlighted the significance of KLF4/MAPK signal interaction in regulating TAM resistance of breast cancer, and suggested that targeting KLF4/MAPK signaling may be a potential therapeutic strategy for breast cancer treatment, especially for the TAM-resistant patients.