Overexpression of NOP58 Facilitates Proliferation, Migration, Invasion, and Stemness of Non-small Cell Lung Cancer by Stabilizing hsa_circ_0001550.

BACKGROUND: NOP58 ribonucleoprotein (NOP58) is associated with the recurrence of lung adenocarcinoma. AIMS: Few investigations concentrate on the role of NOP58 in non-small cell lung cancer (NSCLC), which is the focus of our current study. METHODS: Following transfection, the proliferation, migration, and invasion of NSCLC cells were assessed by 5- ethynyl-2'-deoxyuridine (EdU), wound healing, and transwell assays. The percentage of CD9+ cells was evaluated by flow cytometry assay. Based on target genes and binding sites predicted through bioinformatics analysis, a dual-luciferase reporter assay was performed to verify the targeting relationship between hsa_circ_0001550 and NOP58. The effect of NOP58 overexpression on hsa_circ_0001550 stability was gauged using Actinomycin D. The hsa_circ_0001550 and NOP58 expression levels, as well as protein expressions of CD44, CD133, OCT4, and SOX2 in NSCLC cells were determined by quantitative real-time PCR and Western blot, respectively. RESULTS: Hsa_circ_0001550 was remarkably up-regulated in NSCLC cell lines A549 and PC9, silencing of which weakened cell abilities to proliferate, migrate and invade, decreased CD9+ cell ratio, and diminished protein expressions of CD44, CD133, OCT4, and SOX2. NOP58 could bind to hsa_circ_0001550 and stabilize its expression, and NOP58 overexpression partially abrogated hsa_circ_0001550 knockdown-inhibited NSCLC cell proliferation, migration, invasion and stemness. CONCLUSION: Overexpression of NOP58 facilitates proliferation, migration, invasion, and stemness of NSCLC cells by stabilizing hsa_circ_0001550, hinting that NOP58 is a novel molecular target for NSCLC therapy.

Seismic performance of steel-reinforced concrete-filled rectangular steel tubes after exposure to non-uniform fire.

In actual engineering, non-uniform fire boundary circumstances including single-sided fire, neighboring or related two-sided fire, and three-sided fire, are created due to the varying placements of the columns. In this paper, the seismic performance of SRCFST members subjected to non-uniform fire was investigated by the method of finite element simulation. First of all, the P-Δ curve, ductility coefficient, stiffness, and energy dissipation of the members following non-uniform fire were investigated. As the number of fire surfaces decreases, the maximum overfire temperature at the center of the section decreases, damage decreases, stiffness degradation decreases, and energy dissipation capacity increases. Next, the load distribution of each component in the SRCFST member was calculated using a three-sided fire as an example, the results show that steel tubes play the most dominant role in the seismic performance after fire, followed by steel sections and concrete the least. Last, a parametric study of the key variables influencing the ductility coefficient was carried out.

Analysis of long non-coding RNA expression profile of bovine monocyte-macrophage infected by Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis. As a potential zoonotic pathogen, MAP also seriously threatens human health and social security. At present, long non-coding RNA (lncRNA) has attracted wide attention as an useful biomarker in various diseases. Therefore, our study analyzed the lncRNA expression profiles and lncRNA-mRNA regulatory network of MAP infected bovine monocytes-macrophages and uninfected bovine cells by high-throughput sequencing. A total of 4641 differentially expressed lncRNAs genes were identified, including 3111 up-regulated genes and 1530 down-regulated genes. In addition, lncRNA-mRNA interaction analysis was performed to predict the target genes of lncRNA. Among them, after MAP infection, 86 lncRNAs targeted to mRNA, of which only 6 genes were significantly different. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes significantly enriched in functional groups were related to immune regulation. Multiple signal pathways including NF-κB, NOD-like receptor, Cytokine-cytokine receptor, Toll-like receptor signaling pathway, Chemokine signaling pathway, and other important biochemical, metabolic and signal transduction pathways were enriched in Kyoto Encyclopedia of Genes and Genomes (KEGG). In this study, analysis of macrophage transcriptomes in response to MAP infection is expected to provide key information to deeply understand role of the pathogen in initiating an inappropriate and persistent infection in susceptible hosts and molecular mechanisms that might underlie the early phases of paratuberculosis.

Analysis on Seismic Performance of Steel-Reinforced Concrete-Filled Circular Steel Tubular (SRCFST) Members Subjected to Post-Fire.

Steel-reinforced concrete-filled steel tubular (SRCFST) columns have a great development prospect in engineering practice due to their high load-bearing capacity, good ductility, and energy consumption capacity. This paper established the post-fire seismic analysis model of SRCFST with a circular-cased H section using the sequential coupled thermal-stress method by ABAQUS. The P-Δ curve, stiffness, ductility, and energy dissipation were calculated. Then, the post-fire seismic performance of CFST members was compared while keeping the total steel ratio constant, and it revealed that the SRCFST had superior ductility to CFST. Finally, the ductility coefficient and skeleton curve were parametrically evaluated. The results of the study showed that the effects of heating time (th), axial compression ratio (n), slenderness ratio (λ), and steel tube ratio (αt) on the skeleton line of SRCFST columns are more significant; the axial compression ratio (n), slenderness ratio (λ), and steel tube ratio (αt) have a negative influence on the ductility subjected to post-fire.

Analysis of Fire Resistance of Square-Cased Square Steel Tube Reinforced Concrete (ST-RC) Columns.

Based on the finite element (FE) analysis software Abaqus, an FE model of square-cased square steel tube reinforced concrete (ST-RC) columns under the hybridized action of high-temperature and load is established. The accuracy of the FE model is verified using experimental data from existing studies. This model is used to analyze the temperature change, internal force distribution, and failure characteristics of the square-cased square ST-RC columns under the action of fire, as well as the factors affecting the fire resistance limit of the column. The results of FE analysis show that under the action of fire, the maximum internal temperature of the square-cased square ST-RC columns occurs in the corner of the section. Moreover, the stress and strain reach their maximum values at the concrete corner outside the tube. During the heating process, an internal force redistribution occurs in the square-cased square ST-RC column. At the same time, the proportion of the axial force and the bending moment of the reinforced concrete outside the pipe decreases gradually, while the proportion of the internal force of the core concrete-filled steel tube (CFST) increases gradually. In essence, it is a process of load transfer from the high-temperature to the low-temperature zone. In addition, the section size, load ratio, slenderness ratio, cross-sectional core area ratio, steel content, and external concrete strength are the main parameters affecting the fire resistance limit of the square-cased square ST-RC columns. Among them, the cross-sectional core area ratio, section size, steel ratio, and external concrete strength are positively correlated with the fire resistance limit of the composite column. On the contrary, with the increase in the load ratio and the slenderness ratio, the fire resistance limit of the square-cased square ST-RC columns decreases. On this basis, a simplified formula to calculate the fire resistance limit of square-cased square ST-RC columns is proposed. The research results can be used as a theoretical reference for the fire protection design of this kind of structure in practical engineering.

Small-cell lung cancer transformation from EGFR-mutant adenocarcinoma after EGFR-TKIs resistance: A case report.

RATIONALE: With the recent advancements in molecular biology research, epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have emerged as excellent therapies for patients with EGFR-mutant cancers. However, these patients inevitably develop cross-acquired resistance to EGFR-TKIs. Transformation to small-cell lung cancer (SCLC) is considered a rare resistance mechanism against EGFR-TKI therapy. Here, we report a case of TKI resistance due to SCLC transformation and demonstrate its mechanisms and clinical features. PATIENT CONCERNS: A 54-year-old Chinese man with a history of smoking for 40 years complained of an intermittent cough in March 2019. DIAGNOSIS: Transbronchial lung biopsy was performed on the basal segment of the left lower lobe, which confirmed lung adenocarcinoma. In January 2020, repeat biopsy was performed, and the results of immunohistochemistry (IHC) staining showed TTF-1 (+), CK7 (+), napsin A (+), syn (+), and CD56 (+), with a Ki-67 (+) index 80% of small cell carcinomas. Infiltrating adenocarcinomas and small cell carcinomas were observed. INTERVENTIONS: Icotinib (125 mg thrice daily) was administered as a first-line treatment from June 2019. We subsequently administered a chemotherapy regimen consisting of etoposide (180 mg, days 1-3) plus cisplatin (45 mg, days 1-3) every 3 weeks for 1 cycle after recurrence. As the patient could not tolerate further chemotherapy, he continued taking icotinib orally and received whole-brain radiotherapy 10 times to a total dose of 30 Gy after brain metastases. OUTCOMES: The patient relapsed after successful treatment with icotinib for 9 months. A partial response was achieved after 4 cycles of chemotherapy, and despite the brief success of chemotherapy, our patient exhibited brain metastasis and metastases of the eleventh thoracic spine and the second lumbar vertebra with pathological fracture. The patient eventually died of aggressive cancer progression. LESSONS: Our case highlights the possibility of SCLC transformation from EGFR-mutant adenocarcinoma and the importance of repeat biopsy for drug resistance. Serum neuron-specific enolase levels may also be useful for detecting early SCLC transformation.

[Prospective study of red meat intake in the first and second trimesters and the risk of gestational diabetes mellitus in Chengdu in 2017].

OBJECTIVE: To evaluate the red meat intake of pregnant women in Chengdu area and explore the effect of red meat intake in the first and second trimesters on gestational diabetes mellitus(GDM). METHODS: From February to July 2017, a prospective study was conducted among healthy singleton pregnant women within 8-14 weeks of gestation in a prenatal clinic of maternal-and-child medical institution in Chengdu City through purposive sampling. Data on maternal demographic characteristics was collected through questionnaire in early pregnancy. 3-day 24 hour dietary recall was used to collect dietary intake information in the first and second trimesters. The 75 g oral glucose tolerance test(OGTT)was conducted at 24 to 28 weeks of gestation, and GDM was diagnosed according to the Guidelines for the Diagnosis and Treatment of Pregnancy Diabetes in China(2014). Multivariate unconditional Logistic regression model was used to explore the association between tertiles of red meat intake and the risk of GDM in the first and second trimesters. RESULTS: A total of 985 valid samples were followed up. The mean intake of total meat and red meat in middle pregnancy(102. 9 g/d and 74. 6 g/d) was higher than that in early pregnancy(70. 7 g/d and 52. 0 g/d). The difference was statistically significant(P<0. 05). Red meat accounted for 73. 6% and 72. 5% of total meat intake in early and middle pregnancy, respectively. After adjusting the confounding factors, multivariate unconditional Logistic regression analysis showed the risk of GDM was 1. 499 times(95%CI 1. 028-2. 185) in the highest red meat intake(>86 g/d)women compared with the lowest red meat intake(<44 g/d) women in the second trimester. No association was found between red meat intake in the first trimester and the occurrence of GDM(OR=1. 029, 95%CI 0. 716-1. 481). CONCLUSION: Pregnant women in Chengdu area have excessive intake of red meat in the second trimester. Higher intake of red meat in the second trimester(>86 g/d) may increase the risk of gestational diabetes.

Analysis of mRNA and circRNA Expression Profiles of Bovine Monocyte-Derived Macrophages Infected With Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the pathogen of Johne's disease (paratuberculosis), which mainly causes chronic infectious granulomatous enteritis in ruminants and has brought huge economic losses to animal husbandry. As a specific intracellular pathogen, when MAP invades the body, it is internalized by macrophages where it is able to replicate by inhibition of the phagosome maturation, escaping the host immune system and surviving, which leads to the spread of the disease. More recent studies have shown that circRNA is involved in many pathological and physiological processes of the body as the molecular sponge of miRNA, the scaffold of RNA binding protein and having the characteristic of being able to translate into protein. In this study, the mRNA and circRNA expression profiles of MAP-infected bovine monocyte-macrophages and uninfected bovine cells were analyzed by high-throughput sequencing. A total of 618 differentially expressed mRNA were screened out, including 322 upregulated mRNA and 296 downregulated mRNA. In addition, the analysis of circRNA differential expression profile showed 39 differentially expressed genes including 12 upregulated and 27 downregulated genes. Moreover, differential genes belonging to cytokine activity, chemokine activity, inflammatory reaction, apoptosis, and other functional groups related to macrophage immune response were significantly enriched in Gene Ontology (GO). Multiple signal pathways including NF-κB, MAPK, Toll-like receptor, IL-17, JAK-STAT, and other signaling pathways related to activating macrophage immune response were significantly enriched in Kyoto Encyclopedia of Genes and Genomes (KEGG). In addition, RT-qPCR technology verified the accuracy of the mRNA sequencing results. In this study, we have obtained the transcriptome information of mRNA and circRNA of bovine monocyte-macrophage infected with MAP. These results will provide data support for the further study of mRNA-miRNA-circRNA network and immune escape mechanism of MAP and will enrich the knowledge of the molecular immune mechanisms of Johne's disease as well.

Excessive gestational weight gain in the first trimester is associated with risk of gestational diabetes mellitus: a prospective study from Southwest China.

OBJECTIVE: To evaluate the effects of gestational weight gain (GWG) in the first trimester (GWG-F) and the rate of gestational weight gain in the second trimester (RGWG-S) on gestational diabetes mellitus (GDM), exploring the optimal GWG ranges for the avoidance of GDM in Chinese women. DESIGN: A population-based prospective study was conducted. Gestational weight was measured regularly in every antenatal visit and assessed by the Institute of Medicine (IOM) criteria (2009). GDM was assessed with the 75-g, 2-h oral glucose tolerance test at 24-28 weeks of gestation. Multivariable logistic regression was performed to assess the effects of GWG-F and RGWG-S on GDM, stratified by pre-pregnancy BMI. In each BMI category, the GWG values corresponding to the lowest prevalence of GDM were defined as the optimal GWG range. SETTING: Southwest China. PARTICIPANTS: Pregnant women (n 1910) in 2017. RESULTS: After adjusting for confounders, GWG-F above IOM recommendations increased the risk of GDM (OR; 95 % CI) among underweight (2·500; 1·106, 5·655), normal-weight (1·396; 1·023, 1·906) and overweight/obese women (3·017; 1·118, 8·138) compared with women within IOM recommendations. No significant difference was observed between RGWG-S and GDM (P > 0·05) after adjusting for GWG-F based on the previous model. The optimal GWG-F ranges for the avoidance of GDM were 0·8-1·2, 0·8-1·2 and 0·35-0·70 kg for underweight, normal-weight and overweight/obese women, respectively. CONCLUSIONS: Excessive GWG in the first trimester, rather than the second trimester, is associated with increased risk of GDM regardless of pre-pregnancy BMI. Obstetricians should provide more pre-emptive guidance in achieving adequate GWG-F.

[Energy intake as determinants of gestational weight gain in Chengdu].

OBJECTIVE: To examine the relation between energy intake and its sources and gestational weight gain( GWG). METHODS: In this prospective cohort study, 585 healthy women with singleton pregnancy were selected in 2013. The 24-hour recalls was used to collect food intake at the first, second and last trimesters, and energy intake and percentages of energy from macronutrient were calculated. Weight measuring were administered at( 12 ± 1) th, ( 28 ± 1) th weeks' gestation and last week before delivery, and the GWG rate was calculated. Multivariable linear regression was used to analyze therelationship between energy and GWG. RESULTS: The mean energy intake for the first, second and last trimesters were 1718. 6 kcal/d, 2202. 9 kcal/d and 2313. 4 kcal/d, respectively, the average percentages of energy from carbohydrate were 62. 6%, 59. 5%and 59. 1%, respectively, the average percentages of energy from fat were 21. 8% 、23. 9% and 24. 3%, respectively, the average percentages of energy from protein were15. 6% 、16. 5% and 16. 7%, respectively. Multiple linear regression analysis showed that energy intake during pregnancy was positively correlated with GWG during pregnancy( P < 0. 05). The adjusted weight gain for each trimester was 37. 3 g/week( 95% CI 22. 8 to 51. 8, P < 0. 05), 16. 8 g/week( 95% CI 4. 8 to 28. 8, P < 0. 05), and 25. 8 g/week( 95% CI 10. 6 to 41. 1, P < 0. 05) higher in the highest( Q5) versus lowest( Q1)quintile of energy intake. The adjusted weight gain in last trimester was 19. 1 g/week higher( 95% CI 3. 1 to 35. 1, P < 0. 05) in the highest( Q5) versus lowest( Q1) quintile of percentages of energy from carbohydrate. The adjusted weight gain in last trimester was16. 2 g/week lower( 95% CI-3. 1 to-0. 9, P < 0. 05) in the highest( Q5) versus lowest( Q1) quintile of percentages of energy from fat. CONCLUSION: A positive correlation is found between dietary energy intake and GWG. Percentages of energy from carbohydrate and fat in the last trimester have a significant effect on GWG.

[Effect of gestational weight gain on the neonatal birth weight among pregnancy women in Chengdu City].

OBJECTIVE: To explore the effects of total gestational weight gain( GWG)and gestational weight gain rate( GWGR) per trimester on the neonatal birth weight. METHODS: A prospective cohort study was conducted among 549 healthy pregnant women at the( 12 ± 1) th pregnant week from Chengdu City, China who delivered a singleton. Basic information about age, pre-gestational weight and so on was collected by questionnaire, while the height and weight were measured at the first time. Weight at the( 28 ± 1) th and last week before delivery was measured respectively, while the total GWG and the different trimester GWGR were calculated and assessed by Institute of Medicine( IOM) criteria( 2009). The gestational week, neonatal weight and other information were collected after delivery. Multiple non-conditional Logistic regression models were used to test the effect of the total GWG/GWGR per trimester on neonatal birth weight. RESULTS: The total GWG was( 16. 2 ± 4. 6) kg. The prevalence of abnormal total GWG was 59. 1%, the excessive GWG was 44. 3%. The prevalence of insufficient GWGR in the first trimester was 44. 3%. In the second, third, the second and third trimester, the prevalence of excessive GWGR were 63. 6%, 55. 7% and 65. 8%, respectively. After adjusting the age at delivery, pre-pregnancy body mass index, gestational week and other confounding factors, the result showed that compared with adequate GWG group, insufficient GWG group was increased risk of small for gestational age( SGA)( OR =2. 51, 95% CI 1. 08-5. 82), excessive total GWG group was increased risk of large for gestational age( LGA)( OR = 2. 54, 95% CI 1. 20-5. 36). Compared with adequate GWGR group in the second trimester, excessive GWGR group was decreased risk of SGA( OR = 0. 27, 95% CI 0. 13-0. 60). Compared with adequate GWGR group in the second and third trimester, excessive GWGR group was decreased risk of SGA( OR = 0. 28, 95%CI 0. 13-0. 59). CONCLUSION: Abnormal total GWG among pregnant women in Chengdu City is a big problem that both excessive and insufficient GWG existed. Excessive total GWG is the independent risk factor for LGA. GWGR in the second and the second and third trimester are significantly associated with SGA.

[Effect of gestational weight gain rate on pregnancy outcomes among Chengdu City pregnant women with normal pre-pregnancy body mass index].

OBJECTIVE: To explore the association between gestational weight gain rate( GWGR) and pregnancy outcomes among pregnant women with normal pre-pregnancy body mass index( BMI). METHODS: In this prospective study, 389 healthy pregnant women at the( 12 ± 1) th with normal pre-pregnancy BMI from Chengdu City who delivered a singleton were included in 2013. The basic information such as age, pre-pregnant weight was obtained by questionnaire, height and weight was measured. Then the weight was measured at the( 28 ± 1) th and last week before delivery, respectively, the different trimester GWGR was calculated and classified by IOM criteria. Gestational age, gestational diabetes mellitus( GDM) and other pregnancy outcomes were collected by questionnaire. Multiple non-conditional Logistic regression models were used to test the association between GWGR and pregnancy outcomes. RESULTS: The proportion of insufficient GWGR and excessive GWGR in first trimester was 50. 7% and 31. 8%, respectively, the proportion of excessive GWGR in the second and third trimester was68. 5% and 57. 8%, respectively. After adjusting the age, education, gravidity and family disease history and other confounding factors, multiple non-conditional logistic regression showed: compared with adequate GWGR in the third trimester, insufficient and excessive GWGR were associated with increased risk of cesarean delivery( OR = 2. 48, 95% CI 1. 18-5. 23 and OR = 1. 76, 95% CI 1. 02-3. 03, respectively). Compared with adequate GWGR in the first trimester, excessive GWGR were associated with increased risk of GDM( OR = 2. 55, 95% CI 1. 03-6. 28). CONCLUSION: The abnormal of GWGR would increase the risk of cesarean delivery and GDM.

[Moderate-to-vigorous Physical Activities and Gestational Weight Gains during the Second and Last Trimesters of Pregnancy].

OBJECTIVE: To determine the level of moderate-to-vigorous physical activities (MVPA) and its relationship with gestational weight gains (GWG) in the second and the last trimesters of pregnancy. METHODS: A prospective study was conducted in Chengdu on 362 healthy pregnant women at the 24-28 gestation weeks who delivered a singleton. Demographic data and pre-pregnancy body mass were collected using a questionnaire. Weight gains at the gestation weeks of 24-28 and 32-36 were measured for the first two trimesters and the last trimester of pregnancy. The Denmark self-reported physical activity scale was used for measuring the duration and intensity of physical activities. Multiple linear regression models were established to determine the relationship between MVPA and GWG. RESULTS: The last trimester had lower average daily MVPA 〔(0.76±0.93) h〕 compared with the second trimester 〔(1.61±1.61) h, t=9.056, P<0.001〕. About 74.6% of the participants met the PA recommendations for the second trimester, compared with 60.5% for the last trimester (χ2=16.387, P<0.001). The participants experienced an average GWG of (7.36±3.78) kg during the first two trimesters, and (5.80±2.57) kg during the last trimester, corresponding to a growth rate of (0.30±0.15) kg/week for the first two trimesters and (0.51±0.22) kg/week for the last trimester. Compared with the most inactive group, the participants with medium PA experienced less GWG 〔(5.34±2.91) kg vs.(6.26±2.54) kg, P<0.05〕 and a lower GWG rate 〔(0.48±0.26) kg/week vs.(0.56±0.20) kg/week, P<0.05〕 during the last trimester. Age, gestational week, ethnicity, pre-pregnant BMI, GDM, pre-pregnant smoking and employment were associated with GWG and the GWG rates during the first two trimesters and the third trimester (P<0.05). Compared with the most inactive group, low 〔-0.358(-0.691--0.026)〕 and medium 〔-0.762(-1.486- -0.037)〕 PA were associated with lower GWG during the last trimester. Moderate PA was associated with a lower GWG rate 〔-0.071(-0.133--0.008)〕 after adjustment for gestational age, energy intake, pre-pregnancy BMI and other potential confounders. CONCLUSIONS: Insufficient physical activities are a serious problem in the pregnant women of Chengdu over the last two trimesters. Appropriate MVPA in the last trimester of pregnancy may reduce GWG and GWG rates.

[Effect of microRNA-29b on proliferation and migration of breast cancer cells and its molecular mechanism].

OBJECTIVE: To investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism. METHODS: The recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA. RESULTS: Recombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all P<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all P<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (P<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all P<0.05). CONCLUSIONS: MiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.

Radiosensitization effect of hsa-miR-138-2-3p on human laryngeal cancer stem cells.

BACKGROUND: Treatments that target cancer stem cells play an important role in the controlling and eliminating of tumor initiation as well as in development, progression, and chemotherapy/radiotherapy resistance. In our previous study, we cultured and harvested human laryngeal cancer stem cells (CSCs) and applied microRNA biochips to screen differentially expressed miRNAs that were related to radiation tolerance in irradiated human laryngeal CSCs. According to the predicted genes and pathways of differential miRNAs target, down-regulated expression of hsa-miR-138-2-3p under radiation was thought to play a key role in enhancing the radio-sensitivity in human laryngeal squamous cancer stem cells. METHOD: To investigate the radiational enhancement of hsa-miR-138-2-3p, we transfected hsa-miR-138-2-3p mimics that were synthesized based on the sequences of hsa-miR-138-2-3p in vitrointo human laryngeal CSCs (Hep-2, M2e, and TU212 cell lines) to make hsa-miR-138-2-3p overexpressed, and the tumorous specialities of CSCs, like cell proliferation, invasion, apoptosis, cell cycle arrest, and DNA damage were evaluated by CCK-8 assay, clone formation assay, invasion assay, flow cytometry, and comet assay. Furthermore, we explored the signal transduction pathways that regulated the cancer stem cell initiation, development, invasion, apoptosis and cell cycle arrest, which were controlled by hsa-miR-138-2-3p. RESULT: Overexpressed hsa-miR-138-2-3p played a key role in many anti-cancer biological processes in human laryngeal CSCs: (1) it decreased laryngeal CSCs proliferation and invasion in response to radiotherapy; (2) it increased the proportion of early and late apoptosis in laryngeal CSCs after radiation, raised G1 phase arrest in laryngeal CSCs after radiation, and decreased the proportion of S stage cells of cell cycle that were related to radio-resistance in laryngeal CSCs; (3) it down-regulated the expression of ß-catenin in Wnt signal pathway that was related to the tolerance of laryngeal CSCs to radiotherapy; (4) it down-regulated the expression of YAP1 in Hippo signal pathway that regulated cell proliferation, invasion and apoptosis; (5) it up-regulated the expression of p38 and JNK1 in MAPK signal pathway that was concerned to radio-sensitivity. CONCLUSION: In the present study, it was found that hsa-miR-138-2-3p regulated the Wnt/ß-catenin pathways, the Hippo/YAP1 pathways, and the MAPK/p38/JNK1 pathways that were involved in cell proliferation, invasion, apoptosis, cell cycle arrest, radio-resistance and radio-sensitivity in laryngeal CSCs. These results will be useful for a better understanding of the cell biology of hsa-miR-138-2-3p in laryngeal CSCs, and for serving hsa-miR-138-2-3p as a promising biomarker and as a target for diagnosis and for novel anti-cancer therapies for laryngeal cancers.

Downregulation of caveolin-1 increases the sensitivity of drug-resistant colorectal cancer HCT116 cells to 5-fluorouracil.

Colorectal cancer is the third most common type of cancer in men and women. Chemotherapy is an important treatment strategy for patients with terminal stage cancer. However, the development of drug resistance hampers the effectiveness of chemotherapy. Therefore, an effective therapeutic approach to target chemoresistance-associated cellular molecules is required. In the present study, drug-resistant human colorectal cancer HCT116 cells were developed by treating HCT116 cells with increasing concentrations of 5-fluorouracil (5-FU). The present study indicated that the drug-resistance cells (DRC) were resistant to 5-FU compared with parental HCT116 cells by detecting cell survival using an MTT assay. Additionally, the expression of the chemoresistance-associated protein caveolin-1 (Cav-1) was assessed by reverse transcription-quantitative polymerase chain reaction and western blotting. The results revealed that the Cav-1 expression level was significantly higher in DRC compared with that in the parental HCT116 cells. Next, Cav-1 was silenced by small interfering RNA (siRNA) or was inhibited with its specific inhibitor methyl ß-cyclodextrin (MCD). MTT assay demonstrated that Cav-1 siRNA and MCD resensitized DRC to 5-FU. These data reveal that Cav-1 was involved in the development of resistance, suggesting that Cav-1 is a potential target for the treatment of colorectal cancer chemoresistance. In addition, 5-FU combined with Cav-1 siRNA or its specific inhibitor may increase the effectiveness of the treatment strategy.