MiR-30c facilitates natural killer cell cytotoxicity to lung cancer through targeting GALNT7

Gao Fei,Han Jianjun,Jia Li,He Jun,Wang Yun,Chen Mi,Liu Xiaojun,He Xia 한국유전학회 2023 Genes & Genomics Vol.45 No.2

Background MicroRNAs (miRNAs) have been reported to play important roles in regulating natural killer (NK) cell cytotoxicity to cancer cells. Objective This study aimed to investigate the effects and potential mechanism of miR-30c in regulating NK cell cytotoxicity to lung cancer cells. Methods Primary NK cells were derived from the peripheral blood of lung cancer and normal participants. Exosomes were isolated and validated via transmission electron microscopy and nanoparticle tracking analysis. The levels of miR-30c, polypeptide N-acetylgalactosaminyltransferase 7 (GALNT7) and proteins in PI3K/AKT pathway were determined using quantitative real-time polymerase chain reaction or western blot. Tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) levels and the cytotoxicity of effector NK cells to target lung cancer cells were measured via enzyme linked immunosorbent assay, cell apoptosis or xenograft experiments. The relationship between miR-30c and GALNT7 was analyzed by luciferase activity, RNA pull-down and RNA immunoprecipitation assays. And a xenograft mice model was established to verify the effect of miR-30c in regulating NK cell cytotoxicity to lung cancer cells in vivo. Results NK cell-derived exosomes carrying miR-30c, and miR-30c level was significantly downregulated in primary NK cells of lung cancer patients. MiR-30c overexpression promoted TNF-α and IFN-γ secretion and enhanced the cytotoxicity of interleukin 2 (IL-2)-treated NK cells to lung cancer cells, while knockdown of miR-30c played an opposite effect in regulating the cytotoxicity of NK cells to lung cancer cells. GALNT7 was a target of miR-30c and was negatively regulated by miR-30c. Besides, miR-30c targeted GALNT7 to exert its function in regulating NK cell cytotoxicity. Furthermore, GALNT7 prompted the activation of PI3K/AKT pathway in NK cells. Additionally, miR-30c overexpression enhanced NK cell cytotoxicity to lung cancer cells and inhibited tumor growth in vivo. Conclusion miR-30c enhanced NK cell cytotoxicity to lung cancer cells via decreasing GALNT7 and inactivating the PI3K/AKT pathway, suggesting that regulating miR-30c expression maybe a promising approach for enhancing NK cell-based antitumor therapies.

CircCEP85 upregulates IGF1 expression to promote breast cancer progression via sponging miR‑1193

Gao Fei,Han Jianjun,Jia Li,He Jun,Wang Yun,Chen Mi 한국응용생명화학회 2022 Applied Biological Chemistry (Appl Biol Chem) Vol.65 No.4

Background: Increasing evidence has suggested that circular RNAs (circRNAs) play critical roles in breast cancer (BC) progression. However, the expression level and potential functional role of circRNA centrosomal protein 85 (circ- CEP85) in BC remains largely unknown. Here, we aimed to explore the role of circCEP85 in BC. Methods: The levels of circCEP85, insuline-like growth factor I (IGF1) mRNA and microRNA-1193 (miR-1193) were examined by quantitative real-time polymerase chain reaction. The protein level was measured by Western blot. Cell proliferation, migration, apoptosis, angiogenesis and stemness were assessed by cell counting kit-8, 5-ethynyl- 2’-deoxyuridine assay, transwell assay, flow cytometry, tube formation and sphere formation assays. Xenograft mouse models were conducted to evaluate the effect of circCEP85 in BC in vivo. Moreover, dual-luciferase reporter, RNA pulldown, and RNA immunoprecipitation (RIP) assays were preformed to confirm the interaction between miR-1193 and circCEP85 or IGF1. Results: CircCEP85 was upregulated in BC tissues and cells. Silencing of circCEP85 inhibited proliferation, invasion, angiogenesis and stemness, but promoted apoptosis in BC cells in vitro. In addition, circCEP85 silencing inhibited tumor growth in vivo. Mechanistically, circCEP85 elevated IGF1 expression via sponging miR-1193 to promote breast cancer progression. Conclusion: The circCEP85-miR-1193-IGF1 axis regulated BC progression via the competitive endogenous RNA (ceRNA) mechanism. CircCEP85 might be a prognostic biomarker and therapeutic target for BC.

Physicochemical Characterization and an Injection Formulation Study of Water Insoluble ZCVI4-2, a Novel NO-Donor Anticancer Compound

Yuan Gao,Jianjun Zhang,Li Li,Feng Su,Zhenhua Gong,Yisheng Lai,Yihua Zhang 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.7

ZCVI4-2 was a novel nitric oxide-releasing glycosyl derivative of oleanolic acid that displayed strong cytotoxicity selectively against human hepatocellular carcinoma in vitro and in vivo. In this study, ZCVI4-2 was characterized by FT-IR spectroscopy, differential scanning calorimetry,powder X-ray diffractometry, Raman spectroscopy, hygroscopicity and stability. A high performance liquid chromatography method was also established for the quantitative determination of solubility and additional stability profile of ZCVI4-2. ZCVI4-2 was found to be an amorphous and stable solid with low solubility of less than 10 μg/mL. Based on the solubilization tests that included methods of cosolvency and micellization, the solution mixture of 5% Solutol HS-15, 5% 1, 2-propylene glycol and 5% anhydrous ethanol was determined to be the system for the preparation of the ZCVI4-2 early injection solution. The effect of pH, temperature, light and injectable isotonic glucose or NaCl solution on ZCVI4-2 injection was also investigated. Good stability was observed at all testing conditions. Under the conditions studied, the NO-releasing rate and amount of ZCVI4-2 from the early injection solution in rat plasma demonstrated a promising therapeutic efficacy while maintaining a good safety profile.

BK Knockout by TALEN-Mediated Gene Targeting in Osteoblasts: KCNMA1 Determines the Proliferation and Differentiation of Osteoblasts

Hei, Hongya,Gao, Jianjun,Dong, Jibin,Tao, Jie,Tian, Lulu,Pan, Wanma,Wang, Hongyu,Zhang, Xuemei Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.7

Large conductance calcium-activated potassium (BK) channels participate in many important physiological functions in excitable tissues such as neurons, cardiac and smooth muscles, whereas the knowledge of BK channels in bone tissues and osteoblasts remains elusive. To investigate the role of BK channels in osteoblasts, we used transcription activator-like effector nuclease (TALEN) to establish a BK knockout cell line on rat ROS17/2.8 osteoblast, and detected the proliferation and mineralization of the BK-knockout cells. Our study found that the BKknockout cells significantly decreased the ability of proliferation and mineralization as osteoblasts, compared to the wild type cells. The overall expression of osteoblast differentiation marker genes in the BK-knockout cells was significantly lower than that in wild type osteoblast cells. The BK-knockout osteoblast cell line in our study displays a phenotype decrease in osteoblast function which can mimic the pathological state of osteoblast and thus provide a working cell line as a tool for study of osteoblast function and bone related diseases.

A New Method for Piping Risk Evaluation on Unconfined Aquifers under Dewatering of Deep Foundation Pits

Da Li,Jianjun Ma,Chaosheng Wang,Xiaojuan Gao,Mingjing Fang 대한토목학회 2022 KSCE JOURNAL OF CIVIL ENGINEERING Vol.26 No.8

This paper presents a new method to evaluate the piping risk of unconfined sandy cobble aquifers during the dewatering of deep foundation pits. Firstly, the possibility of piping is judged according to the particle size distribution and pore diameter of the sandy cobble stratum. Secondly, the critical hydraulic gradients of particles with different sizes are calculated according to the pipe flow model. Thirdly, the hydraulic gradient around the foundation pit during the dewatering process is calculated by numerical method. Then, combined with the distribution of hydraulic gradient around the foundation pit and the critical hydraulic gradients of particles with different sizes, the range of moveable particles with different sizes in the surrounding stratum is evaluated. Finally, according to the sandy cobble aquifers conditions and the influence of particle movement on the stratum, the level of piping risk is determined. This method was successfully applied to evaluate the piping risk of Tawan Station in Luoyang Metro Line No.1, and according to the risk evaluation the pensile curtain was recommended for the retaining structure in this case. From the satisfied applications, this method is a reference for the evaluation of piping risk and selection of retaining structure when considering the impact of the dewatering in deep foundation pit with unconfined aquifers on the surrounding stratum.

BK Channel Deficiency in Osteoblasts Reduces Bone Formation via the Wnt/β-Catenin Pathway

Jiang, Lan,Yang, Qianhong,Gao, Jianjun,Yang, Jiahong,He, Jiaqi,Xin, Hong,Zhang, Xuemei Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.8

Global knockout of the BK channel has been proven to affect bone formation; however, whether it directly affects osteoblast differentiation and the mechanism are elusive. In the current study, we further investigated the role of BK channels in bone development and explored whether BK channels impacted the differentiation and proliferation of osteoblasts via the canonical Wnt signaling pathway. Our findings demonstrated that knockout of Kcnma1 disrupted the osteogenesis of osteoblasts and inhibited the stabilization of β-catenin. Western blot analysis showed that the protein levels of Axin1 and USP7 increased when Kcnma1 was deficient. Together, this study confirmed that BK ablation decreased bone mass via the Wnt/β-catenin signaling pathway. Our findings also showed that USP7 might have the ability to stabilize the activity of Axin1, which would increase the degradation of β-catenin in osteoblasts.

Settlement behaviours and control measures of twin-tube curved buildings-crossing shield tunnel

Jianwei Jia,Ruiqi Gao,Defeng Wang,Jianjun Li,Ziwen Song,Jinghui Tan 국제구조공학회 2022 Structural Engineering and Mechanics, An Int'l Jou Vol.84 No.5

Settlement control techniques are critical for the safety of shield tunnel constructions, especially for facing complex situations. In this study, the shield tunnel structure from Huaita east road station to Heping Road station in Xuzhou metro No.3 line (China) is taken as engineering background, which has various complex problems of the upper-soft and lower-hard composite stratum conditions, twin curve shield tunnels, and underpass the foundation of the piled raft. The deformation characteristics of shield tunnelling passing through buildings are explored. Subsequently, comprehensive research methods of numerical simulation and field measurement are adopted to analyzing the effectiveness of settlement control by using the top grouting technique. The results show that the settlement of the buildings has obvious spatial characteristics, and the hysteresis effect can be obviously observed in soil deformation caused by shield construction. Meanwhile, the two shield constructions can cause repeated disturbances, reducing the soil deformation's hysteresis effect. Moreover, the shield tunnel's differential settlement is too large when a single line passes through, and the shield construction of the outer curve can cause more significant disturbance in the tunnel than the inside curve. Notably, the proposed process control parameters and secondary topgrouting method can effectively control the deformation of the shield tunnel, especially for the long-term deformation.

BK Knockout by TALEN-Mediated Gene Targeting in Osteoblasts: KCNMA1 Determines the Proliferation and Differentiation of Osteoblasts

Xuemei Zhang,Hongya Hei,Jianjun Gao,Jibin Dong,Jie Tao,Lulu Tian,Wanma Pan,Hongyu Wang 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.7

Large conductance calcium-activated potassium (BK) channels participate in many important physiological functions in excitable tissues such as neurons, cardiac and smooth muscles, whereas the knowledge of BK channels in bone tissues and osteoblasts remains elusive. To investigate the role of BK channels in osteoblasts, we used transcription activator-like effector nuclease (TALEN) to establish a BK knockout cell line on rat ROS17/2.8 osteoblast, and detected the proliferation and mineralization of the BK-knockout cells. Our study found that the BK-knockout cells significantly decreased the ability of proliferation and mineralization as osteoblasts, compared to the wild type cells. The overall expression of osteoblast differentiation marker genes in the BK-knockout cells was significantly lower than that in wild type osteoblast cells. The BK-knockout osteoblast cell line in our study displays a phenotype decrease in osteoblast function which can mimic the pathological state of osteoblast and thus provide a working cell line as a tool for study of osteoblast function and bone related diseases.

Synthesis of (co-)polyethylene with broad molecular weight distribution by the heterogenous Ziegler–Natta catalysts via one-pot strategy

Zhi Liu,Xinli Zhang,Haibing Huang,Jianjun Yi,Wei Liu,Weijiao Liu,Hongpeng Zhen,Kejing Gao,Mingge Zhang,Wantai Yang,Qigu Huang 한국공업화학회 2012 Journal of Industrial and Engineering Chemistry Vol.18 No.6

A new method was presented for preparing (co-)polyethylene with broad molecular weight distribution (MWD) and high molecular weight employed by the novel heterogeneous Ziegler–Natta catalysts via one-pot strategy. The preparation of these catalysts involved the introduction of alkoxy silane compounds as electron donors. The influences of the electron donors structure, as well as polymerization conditions such as temperature, molar ratio of Al/Ti, ethylene pressure and the concentration of 1-hexene in feed on the polymerization performance for ethylene (co-)polymerization, were investigated. The morphology of the catalyst particles was characterized by SEM and Ti content of these catalysts was characterized by ICP. The GPC results showed that the obtained homopolyethylene and ethylene/1-hexene copolymer had the widest molecular weight distribution up to ca 50 when diethoxy-isopropoxy-(t–butoxy)-silane (ED3) was used as internal electron donor. The 13C NMR and FT-IR analysis indicated that these catalysts efficiently catalyzed the copolymerization of ethylene with 1-hexene.

Corrosion Resistance and Compressive Strength of Cemented Soil Mixed with Nano-Silica in Simulated Seawater Environment

Qing-Sheng Chen,Hongyu Zhang,Jianjun Ye,Gao-LiangTao,Sanjay Nimbalkar 대한토목학회 2023 KSCE Journal of Civil Engineering Vol.27 No.4

Cemented soil structures are frequently exposed to the corrosive environment of seawater for an extended period, severely affecting their interior structure and mechanical qualities. This study presents laboratory-based approach to investigate the effect of nano-SiO2 on the structural and mechanical properties of cemented soil in a simulated seawater environment. The unconfined compressive strength (UCS), elastic modulus and other mechanical properties of cemented soil mixed with nano-SiO2 and ordinary silicate cemented soil were evaluated. X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to deduce the crystalline composition and microstructure characteristics of the modified cemented soil. The results show that the addition of nano-SiO2 greatly increased the compressive strength and corrosion resistance of the cemented soil. In a 3C (105% salinity) simulated saltwater curing environment, the compressive strength of the cemented soil containing 2% nano-SiO2increased by 86% and 158% at 30days and 60days, respectively. XRD and SEM showed that nano-SiO2 enhanced the interface structure of cemented soil and increased the compactness of the cement-soil system. The current study demonstrates that nano-SiO2 could considerably improve the mechanical characteristics and corrosion resistance of cemented soil when exposed to simulated seawater.