[Expression of ProEXC and PRMT5 in cervical adenocarcinoma and their clinical significance].

This study observed the expression of ProEXC protein and PRMT5 protein in cervical adenocarcinoma and adjacent tissues, exploring the relationship between the expression of ProEXC and PRMT5 and the auxiliary diagnosis of cervical adenocarcinoma, as well as the clinical pathological parameters. A total of 88 specimens diagnosed with cervical adenocarcinoma from the Second Affiliated Hospital of Soochow University between 2015 and 2020 were collected. Immunohistochemistry was employed to detect the expression of ProEXC and PRMT5 in cervical adenocarcinoma and adjacent tissues, and statistical analysis was conducted. The Cancer Genome Atlas (TCGA) database was utilized to analyze the correlation between the prognosis of cervical adenocarcinoma patients and the expression of ProEXC and PRMT5, as well as their related gene pathways. The GSE39293 dataset from the Gene Expression Omnibus (GEO) was selected to compare the expression levels of ProEXC and PRMT5 in cervical adenocarcinoma cell lines (HELA) before and after antiviral drug treatment.In cervical adenocarcinoma tissues, the expression of ProEXC protein (95.5% vs 4.6%, P<0.001) and PRMT5 protein (81.8% vs 26.1%, P<0.001) was significantly higher than in surrounding adjacent tissues. Their expression was correlated with the tumor's T stage, lymph node metastasis, and human papillomavirus (HPV) infection (P<0.05). TCGA database analysis showed that patients with high expression of MCM2 in PRMT5 and ProEXC had a lower overall survival rate (P<0.05), while the expression of TOP2A was not significantly correlated with survival. In the GSE39293 dataset, the expression of MCM2 (9.34 vs 9.68, P<0.001) and PRMT5 (8.16 vs 8.26, P=0.087) in cells decreased after treatment with cidofovir, while TOP2A (8.54 vs 8.42, P=0.056) expression did not change significantly. In the drug-resistant group, the expression of PRMT5 (8.42 vs 8.16, P=0.002) and MCM2 (9.51 vs 9.34, P=0.029) increased, while TOP2A (8.06 vs 8.54, P<0.001) expression decreased. Gene set enrichment analysis (GSEA) suggested that high expression of ProEXC mainly affected the cell cycle pathway, while high expression of PRMT5 mainly affected the RNA splicing pathway.This study found that ProEXC protein and PRMT5 protein were highly expressed in cervical adenocarcinoma tissues, and the high-expression group had a poorer prognosis, showing a certain correlation with the clinical and pathological characteristics of cervical adenocarcinoma. This may be related to their influence on the cell cycle and RNA synthesis pathways, suggesting their potential significant roles in the progression of cervical adenocarcinoma.

[Study on formulation and revision of standard limits for radon in "Standards for indoor air quality (GB/T 18883-2022)" in China].

Public exposure to radon has attracted increasing public concern. The newly issued "Standards for indoor air quality (GB/T 18883-2022)" has revised the radiological parameters of radon. This study analyzed and discussed the relevant technical contents about the derivation of radon limit, including the distribution level for indoor radon, exposure pathway, health effects, and the process for establishing the standard limits. Specific implementation and evaluation suggestions are also proposed.

Comparative analysis of differentially expressed genes related to triglyceride metabolism between intramuscular fat and abdominal fat in broilers.

1. Lipid metabolism is an indispensable process in an organism, though little is known about the regulatory mechanisms of fat deposition in different types of adipose tissues. 2. The differentially expressed genes related to triglyceride (TG) metabolism between abdominal and intramuscular fat (IMF) of Beijing-You chickens were investigated in this study. 3. TG content in abdominal fat (AF) (349.7 mg/g) was significantly higher (P < 0.01) than in the breast and thigh (12.3 mg/g and 24.8 mg/g, respectively). 4. Using Agilent chicken gene-expression profiling in adipose tissues between AF and muscle (breast and thigh), certain representative genes related to fatty acid metabolism, lipoprotein catabolism and esterification reactions were significantly upregulated (P < 0.05 or P < 0.01). 5. Genes involved in fatty acid oxidation or carbohydrate utilisation were significantly up- or downregulated (P < 0.05 or P < 0.01), including those involved with highly enriched pathways of lipid metabolism (PPAR, Wnt pathway and inositol phosphate metabolism), cell junctions (focal adhesion and regulation of actin cytoskeleton) and muscle contraction. 6. Overall, higher TG levels were observed in AF tissue than in adipose tissues of breast and thigh, which could be regulated through gene expression of pathways related to lipid metabolism (PPAR, Wnt pathway and inositol phosphate metabolism), cell junctions (focal adhesion and regulation of actin cytoskeleton) and muscle contraction. These results provide clues to understanding the molecular mechanisms of TG metabolism between abdominal and IMF.

Effect of 1,25-(OH)2D3 and lipopolysaccharide on mononuclear cell inflammation in type 2 diabetes mellitus and diabetic nephropathy uremia.

The prevention and treatment of type-2 diabetes mellitus (T2DM) and diabetic nephropathy (DN), which are disorders with high incidence rates, is of primary importance. In this study, we analyzed the effect of 1,25-(OH)2D3 and lipopolysaccharide (LPS) in combination with interleukin (IL)-15 on the inflammatory immune response and expression of vitamin D receptor (VDR) in mononuclear cells of T2DM and DN uremia (DNU) patients. The human acute monocytic leukemia cell line THP-1 was treated with peripheral blood serum isolated from 30 healthy controls and T2DM and DNU patients each, cultured in the presence or absence of 1,25-(OH)2D3, and subsequently treated with LPS and IL-15. The VDR mRNA and protein expression in THP-1 cells was detected by real-time polymerase chain reaction and western blot (and immunofluorescence assay), respectively, and IL-6 and IL-10 concentrations in the culture supernatant were detected by enzyme-linked immunosorbent assay. LPS treatment induced a significant decrease in VDR mRNA expression in T2DM and DNU serum-treated THP-1 cells compared to the control cells (P < 0.05). The VDR protein expression in DNU serum-treated THP-1 cells was also significantly down-regulated (P < 0.05). LPS treatment induced IL-6 secretion in serum-treated THP-1 cells (P < 0.05), while 1,25-(OH)2D3 treatment inhibited IL-6 secretion to some extent. These findings suggested that LPS down-regulates the expression of VDR in mononuclear cells of T2DM and DNU patients and induces an imbalance in the pro-inflammatory and anti-inflammatory cytokine response, while 1,25-(OH)2D3 partially reversed the effect of LPS and protected patients with T2DM and DNU.

[Efficiency and adverse effects of the effective therapy applying etoposide + cisplatin and its subsequent maintenance therapy with different durations in patients with small cell lung cancer].

OBJECTIVE: To explore the efficiency and adverse effects of the effective EP (etoposide + cisplatin) therapy and its subsequent maintenance therapy with different durations in patients with small cell lung cancer (SCLC). METHODS: Clinical data of 104 SCLC patients diagnosed and treated at the Jilin Province Cancer Hospital between September 2010 and December 2013 were retrospectively analyzed.Among them, 35 patients were subsequently treated with a 4-week maintenance therapy following the original therapeutic regimen after the effective EP therapy (4-week maintenance therapy group), 35 patients were treated with a subsequent 6-week maintenance therapy (6-week maintenance therapy group), and 34 patients were treated without maintenance therapy (control group).52 patients were in limited stage, and 52 patients were in extensive stage. The progression-free survival (PFS), overall survival (OS) and adverse effects in the 4-week maintenance therapy group, 6-week maintenance therapy group and control group were analyzed. RESULTS: The median PFS in the control group, 4-week maintenance therapy group and 6-week maintenance therapy group was 4.0, 3.5, and 4.0 months, respectively, and the median OS was 9.0, 10.0 and 12.0 months, respectively, showing no significant difference among the groups (P>0.05 for all). The median PFS was prolonged by 2 months as compared with the control group after the 4-week maintenance therapy in the patients with complete remission in first-line chemotherapy (P=0.041), while the median OS was not improved (P=0.131). Neither the median PFS nor median OS showed statistically significant difference between each two groups in the patients with partial remission in first-line chemotherapy (P>0.05 for all). In the limited stage, the median PFS in the control group, 4-week maintenance therapy group, and 6-week maintenance therapy group was 5.0, 6.5, and 4.0 months, respectively, and median OS was 11.0, 13.5, and 13.0 months, respectively, the differences showed no statistical significance (P>0.05 for all). In the extensive stage, the median PFS in the control group, 4-week maintenance therapy group, and 6-week maintenance therapy group was 3.0, 3.0, and 3.5 months, respectively, showing significant differences (P=0.015); the median OS was 6.5, 8.0, and 8.0 months, respectively, presenting no statistically significant differences (P=0.096). In addition, the PFS in the 6-week maintenance therapy group was significantly improved as compared with that in the control group (P=0.016). Compared with the control group, the incidence rates of nausea (grade 3-4), vomiting, hypodynamia, leukopenia, neutropenia, and thrombocytopenia in the 4-week maintenance therapy group and 6-week maintenance therapy group were increased significantly (P<0.05 for all), however, the side effects were tolerable. CONCLUSION: Prolonging the treatment cycle of EP therapy can improve the PFS in SCLC patients in first-line CR chemotherapy and extensive stage.

[Comparison of clinical outcomes of patients with non-small cell lung cancer harboring different types of epidermal growth factor receptor sensitive mutations after first-line EGFR-TKI treatment].

OBJECTIVE: To analyze whether there are differences in the efficacy and clinical outcomes to first-line tyrosine kinase inhibitors (TKI) therapy in Chinese patients with metastatic non-small-cell lung cancer (NSCLC) harboring different subtypes of epidermal growth factor receptor (EGFR) mutations. METHODS: A retrospective analysis was made on the clinical data of stage ⅢB or Ⅳ NSCLC patients who were diagnosed by histology and received EGFR mutation test, in order to confirm if there is any difference between the therapeutic effects of TKIs as first-line therapy and the prognosis. RESULTS: A total of 165 patients harboring EGFR exon 19 deletion (19del, n=71), exon 21 L858R mutation (L858R, n=80) or uncommon sensitive mutation (n=14) were treated with EGFR-TKIs for first-line treatment. The comparison among different groups of common types of sensitive mutations revealed that the objective response rate (ORR) of group 19del and group L858R were 57.8% and 45.0%, respectively (P=0.113). The disease control rate (DCR) was 93.0% and 93.8%, respectively (P=0.158). However, the ORR and DCR of uncommon sensitive mutation were 35.7% and 78.6%, which were significantly lower than that of the group 19del (P=0.035) and group L858R (P=0.020). The median progression-free survival (PFS) of group 19del, group L858R and uncommon sensitive mutation were 14.0 months, 7.8 months and 5.1 months, respectively (P=0.001). The median PFS of the group 19del was significantly longer than that of the group L858R (P=0.009). The median overall survival (OS) of these three groups had significant difference (22.8, 15.2 and 10.0 months) (P=0.048). But those of group 19del and group L858R were similar (P=0.152). The multivariate analysis indicated that ECOG-PS (P=0.030), cigarette smoking (P=0.013) and EGFR mutation types (P=0.034) are independent prognostic factors of OS. CONCLUSIONS: For Chinese NSCLC patients with different types of sensitive mutation, there are differences between their efficacy and prognosis of EGFR-TKIs as first-line treatment. The PFS of group 19del is obviously longer than that of other types of sensitive mutations, but have no significant differences in OS. The PFS and OS of patients with common types of sensitive mutation are better than those with uncommon sensitive mutation.

Biocompatibility of differently proportioned HA/PLGA/BMP-2 composite biomaterials in rabbits.

The aim of this study was to assess the histocompatibility of hydroxyapatite (HA)/poly(lactic-co-glycolic acid) (PLGA)/bone morphogenetic protein-2 (BMP-2) composite materials in rabbits. Thirty healthy New Zealand white rabbits were randomly divided into 3 groups (N = 10). HA/PLGA/BMP-2 composite materials with the HA/PLGA proportions of 1:1, 1:2, and 1:3 were implanted in the animals, which were subsequently sacrificed on the 30th and 60th days post-implantation to allow for differences in routine blood and biochemical indices to be assessed between the animal groups. The degree of biomaterial degradation was also assessed in the three groups. Thirty and 60 days after the implantation of titanium plates and composite materials, no rabbits succumbed to inflammatory reactions, adverse reactions, abnormal blood routine and biochemical indices, or unstable liver functions. The presence of newborn tissues was identified within the 60 days post-implantation. No significant differences were observed between the three groups (P < 0.05). The wide clinical application of HA/PLGA/BMP-2 composite biomaterial, which is highly compatible with rabbits with no apparent effects on the animals, is highly feasible.

Correlation between transcription factor activator protein-2ß (TFAP-2ß) and endometrial carcinoma.

AIMS: To investigate the correlation between transcription factor activator protein-2ß (TFAP-2ß) and endometrial carcinoma (EC). MATERIALS AND METHODS: The study comprised 60 randomly selected patients diagnosed and treated at the 2nd Affiliated Hospital of Harbin Medical University from November 2011 to June 2012 for endometrial carcinoma (n = 30) and myoma of uterus (n = 30). The expression of TFAP-2Pß mRNA in endometrial carcinoma was analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). Body mass index (BMI), waist circumference, and venous blood samples were obtained before abdominal surgery clinically. RESULTS: The expression of TFAP-2ß mRNA in endometrial tissue of patients with EC was higher than that of normal endometrium (p < 0.05). The expression of TFAP-2ß mRNA in endometrial tissue of patients with metabolism syndrome was higher than that of lean ones (p < 0.05). There was no significant difference in the expression of TFAP-2ß mRNA in endometrial tissue between patients with both EC and metabolism syndrome and in those with EC only. The expression levels of TFAP-2ß mRNA had positive correlation with triglyceride (r = 0.271, p < 0.05) and high-density lipoprotein (HDL) (r = 0.314, p < 0.05). There was no significant correlation between the expression of TFAP-2ß mRNA and CA125, fasting plasma glucose, low-density lipoprotein (LDL), waist circumference, total cholesterol, and BMI. CONCLUSIONS: TFAP-2ß constituted promoter activity in EC and also contributed to the development of the metabolic syndrome. TFAP-2ß may influence the oc- currence and development of EC through regulating the expression of various adipokines and lipoprotein metabolism. Probably TFAP-2ß can be a candidate tumor marker for EC.

Liver dominant expression of fatty acid synthase (FAS) gene in two chicken breeds during intramuscular-fat development.

Fatty acid synthase (FAS) is a key enzyme of lipogenesis. In this study, the FAS mRNA expression patterns were examined in three fat related tissues (liver, breast and thigh) at different developmental stages in two chicken breeds (Beijing-You, BJY and Arbor Acres broiler, AA). Results of the Real time-qPCR showed that the expression of FAS mRNA level in liver was significantly higher (P < 0.01) than that in breast and thigh in both two chicken breeds. Significant differences of FAS mRNA expression in liver were found between BJY and AA chickens during different developmental stages. After the contents of intramuscular-fat (IMF) and the liver fat were measured, the correlation analysis was performed. In liver, the FAS mRNA level was highly correlated with hepatic fat content (r = 0.891, P < 0.01 for BJY; r = 0.901, P < 0.01 for AA). On the contrary, the FAS expression level in both breast and thigh tissues were relatively low, stable and there was no correlation between the FAS mRNA level and IMF content in breast and thigh tissues of each breed. The results here can contribute to the knowledge on the developmental expression pattern of FAS mRNA and facilitate the further research on the molecular mechanism underlying IMF deposition in chicken.

Comparison of breast muscle meat quality in 2 broiler breeds.

On the basis of meat quality traits, muscle fiber characteristics, and nutrient components and contents in chickens at market age, 120-d-old Beijing-you (BJY) chickens (the Chinese local breed) had distinct breast muscle features when compared with 42-d-old Arbor Acres (AA) chickens (the genetically improved broiler line). The phospholipid (P < 0.05) and essential fatty acid (P < 0.05) contents in BJY chickens were significantly higher than those in AA chickens. No differences (P > 0.05) were found between the breeds in the contents of polyunsaturated fatty acids, unsaturated fatty acids, protein, or amino acids. Breast muscle fiber diameter was significantly smaller (~55.76%) and fiber density was higher (~174.86%) in BJY chickens than in AA chickens (P < 0.05). In this study, breast muscle from 120-d-old BJY chickens was judged to have better quality of phospholipids and essential fatty acid contents and muscle fiber characteristics than breast muscle from 42-d-old AA chickens.

The effect of a mutation in the 3-UTR region of the HMGCR gene on cholesterol in Beijing-you chickens.

The 3-hydroxyl-3-methylglutaryl Coenzyme A reductase (HMGCR) gene was examined for polymorphisms in Beijing-you chickens. A "T" base insert was detected at nucleotide 2749 of the 3-UTR region of the HMGCR gene and was used as the basis for distinguishing a B allele, distinct from the A. Serum and muscle contents of total cholesterol. LDL-cholesterol in serum was significantly lower in AB birds and lowest in BB birds. Real-time PCR showed that the same trends across genotypes occurred in an abundance of HMGCR transcripts in liver, but there was no difference in contents of HMGCR mRNA in breast or thigh muscles. Hepatic expression and serum LDL-cholesterol were meaningfully correlated (partial, with total serum cholesterol held constant, r = 0.923). In muscle, similar genotypic differences were found for the abundance of the LDL receptor (LDLR) transcript. Cholesterol content in breast muscle related to LDLR expression (partial correlation with serum LDL-cholesterol held constant, r = 0.719); the equivalent partial correlation in thigh muscle was not significant. The results indicated that the B allele significantly reduces hepatic abundance of HMGCR transcripts, probably accounting for genotypic differences in serum cholesterol. In muscle, the cholesterol content appeared to reflect differences in LDLR expression with apparent mechanistic differences between breast and thigh.

Long non-coding RNA MNX1-AS1 promoted osteosarcoma proliferation and invasion via inhibiting KISS1.

OBJECTIVE: Recent studies revealed that long non-coding RNAs (lncRNAs) participate in the progression and development of many human diseases. In this work, we are committed to uncovering the association between lncRNA MNX1-AS1 and the development of osteosarcoma. PATIENTS AND METHODS: MNX1-AS1 expression of osteosarcoma cells and tissue samples was detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Besides, we conducted functional assays including cell proliferation assay, colony formation, and transwell assays. Furthermore, the underlying mechanism including RT-qPCR and Western blot assay was performed. RESULTS: MNX1-AS1 was higher-expressed in osteosarcoma samples than that in adjacent tissues. The abilities of proliferation and invasion were suppressed after MNX1-AS1 was knocked down in vitro. Moreover, KISS1 expression was upregulated at mRNA and protein level via silence of MNX1-AS1. Furthermore, the underlying mechanism of the development of osteosarcoma were investigated by RT-qPCR and Western blot assay. CONCLUSIONS: Our study demonstrated that MNX1-AS1 could enhance osteosarcoma cell proliferation and invasion by inhibiting KISS1, which might contribute to the therapy for osteosarcoma.

A selection method of chickens with blue-eggshell and dwarf traits by molecular marker-assisted selection.

The blue-eggshell and dwarf traits have an important economic value in poultry production. Using a genetic aggregation-based strategy, the molecular marker-assisted selection technology was jointly used to provide a rapid breeding method for pure strain chickens simultaneously with hens exhibiting the blue-eggshell and dwarf traits. Overall, 80 male dwarf chickens and 1,000 hybrid blue-eggshell hens (F0) were used for the hybridization experiment. Subsequently, the crossing of F1 or F2 chicks was performed in succession. The F1 and F2 chicks were respectively detected by the joint molecular markers of the solute carrier organic anion transporter family, namely, 1B3 (SLCO1B3) and the growth hormone receptor (GHR) genes, which relate to blue-eggshell and dwarf traits. Meanwhile, the selection of blue-eggshell and dwarf phenotypes was used to validate the data obtained by the molecular markers. The results showed that F1 chicks included the heterozygous and wild-type of SLCO1B3, as well as the homozygous (hens) and heterozygous (roosters) of GHR. However, F2 chicks included 3 different genotypes of both SLCO1B3 and GHR. Ultimately, 196 F1 roosters (concurrently with heterozygous genotype of SLCO1B3 and GHR) and 1,073 F1 hens (concurrently with heterozygous genotype of SLCO1B3 and homozygous genotype of GHR) were obtained from the initial 10,040 F1 chicks. Further, 27 F2 roosters and 345 F2 hens, which simultaneously carried the homozygous genotype of SLCO1B3 and GHR, were screened from the initial 6,000 F2 chicks. Data obtained on the blue-eggshell and dwarf phenotypes were consistent with the results by molecular markers. Similarly, the purity verification of the strain obtained through 2 crossing experiments (F0♂ × F2♀ and F2♂ × F2♀) revealed that all chickens had the blue-eggshell and dwarf traits, supporting that the obtained F2 strain was pure. In summary, for the first time, we successfully bred a pure strain chicken with blue-eggshell and dwarf traits by jointly using the molecular markers of the SLCO1B3 and GHR genes. Our study provides a new method for the rapid cultivation of new chicken strains.

LncRNA SNHG15 promotes proliferation and migration of lung cancer via targeting microRNA-211-3p.

OBJECTIVE: To investigate whether lncRNA (long non-coding RNA) SNHG15 could regulate the proliferation and migration of lung cancer via microRNA-211-3p and its underlying mechanism. PATIENTS AND METHODS: SNHG15 expression in 55 LC (lung cancer) tissues and 30 normal lung tissues was detected by qRT-PCR (quantitative Real Time-Polymerase Chain Reaction). The relationship between SNHG15 expression and pathological characteristics of LC patients was analyzed the by Kaplan-Meier method. The target microRNA of SNHG15 was predicted by bioinformatics and verified by dual-luciferase reporter gene assay. Viability, cell cycle and migration of LC cells after altering expressions of SNHG15 or microRNA-211-3p were detected by cell counting kit-8 (CCK-8), flow cytometry and transwell assay, respectively. RESULTS: SNHG15 was highly expressed in LC tissues than that of normal lung tissues. Besides, LC patients with stage I-II presented lower expression of SNHG15 than those with stage III-IV. SNHG15 expression was correlated to tumor size, TNM stage, and lymph node metastasis, whereas not correlated to age and sex of LC patients. For in vitro studies, SNHG15 knockdown resulted in viability reduction, cell cycle arrest and reduced migration of LC cells, which were reversed by the microRNA-211-3p knockdown. CONCLUSIONS: SNHG15 is highly expressed in LC tissues, which promotes the occurrence and progression of LC via regulating proliferation and migration of LC cells by targeting microRNA-211-3p.

miRNA211 induces apoptosis of cervical cancer SiHa cells via down-regulation of inhibitor of apoptosis proteins.

OBJECTIVE: Cervical cancer severely threatens patients' lives. MicroRNAs contribute to regulatory function in the growth and apoptosis of cells. The present study investigated the effect of miRNA211 on growth and apoptosis of cervical cancer SiHa cells. MATERIALS AND METHODS: miRNA211 and control miRNA were synthesized and transfected into cervical cancer SiHa cells. MTT assay and flow cytometry were used to study the effect of miRNA211 on growth and apoptosis of SiHa cells. RT-PCR and Western blot were used to detect the expression of inhibitor of apoptosis proteins (IAPs) at both mRNA and protein levels. Groups of miRNA (NC), miRNA211, miRNA+siRNA IAP, miRNA211+siRNA IAP were established and levels of IAP and caspase 3 from each group were measured after transfection. RESULTS: After transfection with miRNA211, the growth of SiHa cells was significantly inhibited and apoptosis of SiHa cells was induced, with the reduction of IAPs at both mRNA and protein levels (p<0.05). Knockdown of IAPs enhanced the apoptosis of SiHa cells that were induced by miRNA211, while the overexpression of limited the pro-apoptotic effect of miRNA211 on SiHa cells. CONCLUSIONS: MiRNA211 inhibits the growth of SiHa cells via down-regulation of IAPs.

Long non-coding RNA BANCR indicates poor prognosis for breast cancer and promotes cell proliferation and invasion.

OBJECTIVE: We investigated the expression of human long non-coding ribonucleic acid (lncRNA), BRAF-activated non-coding RNA (BANCR) in breast cancer tissues and its effects on the in vitro proliferation, apoptosis, invasion and metastasis of breast cancer cells; also, we investigated its possible mechanism. PATIENTS AND METHODS: The expressions of BANCR in 65 pairs of breast cancer tissues and para-carcinoma normal breast tissues were detected by Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The expressions of BANCR in normal breast epithelial cells (MCF10A) and breast cancer cells (MCF-7, MDA-MB-231, SKBR3 and BT-20) were further detected. The knockdown efficiency of BANCR small interfering RNA (siRNA) in MCF-7 cells was detected by qRT-PCR. The effects of BANCR knockdown on proliferation, apoptosis, invasion and metastasis capacities of MCF-7 cells were explored via methyl thiazolyl tetrazolium (MTT) proliferation assay, cell colony formation assay, fluorescence-activated cell sorting (FACS) and transwell migration assay. Western blotting was used to detect the changes in expressions of apoptosis-related proteins, epithelial-mesenchymal transition (EMT)-related proteins and matrix metalloproteinases (MMPs) after knockdown of BANCR. RESULTS: The expression level of lncRNA BANCR in breast cancer tissues was significantly higher than that in para-carcinoma normal tissues. The prognosis of patients in low-expression BANCR group was significantly superior to that of patients in high-expression BANCR group. After BANCR knockdown in breast cancer MCF-7 cells, the cell proliferation and colony formation capacities were significantly inhibited. Further mechanism research showed that inhibiting BANCR could promote the apoptosis of MCF-7 cells. Results of Western blotting revealed that the expressions of B-cell lymphoma 2 associated X protein (BAX), cleaved-Caspase-3 and cleaved-poly adenosine diphosphate-ribose polymerase (PARP) in knockdown group were significantly up-regulated compared with those in control group. Both wound-healing assay and transwell migration assay showed that the down-regulation of lncRNA BANCR could inhibit the invasion and metastasis capacities of MCF-7 cells, whose mechanism was related to the inhibition of EMT process and down-regulation of MMP expressions in cells. CONCLUSIONS: LncRNA BANCR is highly expressed in breast cancer, which is significantly correlated with the prognosis of patients; moreover, it can promote the growth, invasion and metastasis of ovarian cancer cells. The down-regulation of BANCR can inhibit the proliferation, invasion and metastasis capacities of MCF-7 cells.

Long non-coding RNA FENDRR inhibits NSCLC cell growth and aggressiveness by sponging miR-761.

OBJECTIVE: The aim of this paper is to investigate the functions of long noncoding RNA (lncRNA) FOXF1 Adjacent Non-Coding Developmental Regulatory RNA (FENDRR) in the growth and aggressiveness of non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: The expression of FENDRR in NSCLC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays were conducted to explore the roles of FENDRR on the growth of NSCLC cell. The wound healing and transwell invasion assays were conducted to explore the impact of FENDRR on NSCLC cell migration and invasion. The apoptosis of NSCLC cell was detected using flow cytometer-based Annexin V/Propidium Iodide (PI) dual staining. The xenograft model was conducted to investigate the effect of FENDRR on the growth of NSCLC cell in vivo. The expression of Ki67 was measured by immunohistochemical (IHC) staining using Ki67 antibody. Bioinformatics analysis and Luciferase reporter assay were applied to identify that miR-761 was the target of FENDRR. Additional, colony formation and transwell experiments were utilized to confirm that FENDRR inhibited the growth and aggressiveness of NSCLC cell by regulating miR-761. RESULTS: We found a marked down-regulation of FENDRR in NSCLC tissues compared to tumor-adjacent tissues. FENDRR down-expression was detected in four NSCLC cell lines (H1650, HCC827, H1975 and A549) compared to the human non-tumorigenic bronchial epithelial cell, BEAS-2B. Low expression of FENDRR was identified as a predictive factor for poor prognosis of patients with NSCLC. The over-regulation of FENDRR inhibited the proliferation, migration and invasion capacities of NSCLC cell and promoted the apoptosis of NSCLC cell in vitro whereas the down-regulation of FENDRR caused the opposite results. Moreover, the over-expression of FENDRR restrained the growth of NSCLC cell in vivo. We found that there were potential binding sites between FENDRR and miR-761 and the level of miR-761 was inversely associated with the expression of ENDRR in NSCLC tissues. Finally, the rescue experiments suggested that the anti-oncogenic role of FENDRR was at least partially mediated by miR-761 in NSCLC. CONCLUSIONS: We found that FENDRR was down-expressed in NSCLC and the over-expression of FENDRR inhibited the malignant phenotypes of NSCLC cell by binding to miR-761 competitively.

Method using a co-culture system with high-purity intramuscular preadipocytes and satellite cells from chicken pectoralis major muscle.

Intramuscular fat is important in improving meat quality; however, the lack of high-purity intramuscular preadipocyte (IMP) in vitro has severely limited the in-depth research on the mutual regulation of myocytes and adipocytes in chicken. In this study, we establish a new method by combining the mature adipocyte ceiling method and the transwell co-culture system. Mature intramuscular adipocyte (MIA) and muscle satellite cell (MSC) were obtained from digested pectoralis major, and MIAs were transformed into IMPs by dedifferentiation with ceiling culture. MSCs were then purified by differential adhesion for 2 h. The results by inverted-microscope observation, MTT assay, Oil Red O staining, and q-PCR revealed that the de-differentiated cells from MIA were identified as the IMPs, and had the same the cellular morphology, the capacity on differentiation, proliferation and passage with the abdominal fat preadipocytes (P > 0.05). The applicability of the obtained IMPs in co-cultured experiment with the MSC revealed that it could meet the requirements of the experimental study. Finally, a co-culture system of IMPs and MSCs was established using a transwell chamber. The co-cultured results indicated that MSCs in the proliferative stage tend to accelerate the differentiation of IMPs to induce more fat content in co-cultured IMPs than in single-culture IMPs (P < 0.05), in the non-proliferative stage, the results tend to show the opposite (P < 0.05). The mRNA levels of related genes significantly changed in accordance with the fat content in cells. The results strongly supported the view that the established co-culture system was effective and feasible. In summary, we successfully found a new method to explore the interaction between myocytes and adipocytes of chicken. Our findings can deepen research on the regulation of chicken myocytes and adipocytes.