甜瓜质地差异及相关细胞壁酶活性变化

对两种不同质地类型甜瓜果实发育过程中果肉质地的变化及相关酶活性进行研究。结果表明,花后35~40 d为软、脆两种甜瓜质地形成的关键时期。在该时期,软肉型甜瓜‘NSL’细胞面积与间隙增大,果肉细胞排列疏松,细胞面积为脆肉型甜瓜的115.35%,软肉型甜瓜‘NSL’与脆肉型甜瓜‘XZM’质构参数差异显著,脆肉型甜瓜果实4种细胞壁酶活性总体低于软肉型甜瓜。花后35 d,软肉型甜瓜‘NSL’细胞壁扩展酶基因(CmEXP3、CmEXP5、CmEXP9)相对表达量为最大值,显著高于脆肉型甜瓜‘XZM’。     

FAM3C activates Akt to promote viability of oral squamous-cell carcinoma cells

AIM: To investigate the expression and roles of family with sequence similarity 3, member C (FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte (DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with siFAM3C or FAM3C antibody. After 24, 48 and 72 h, the viability of WSU-HN6 cells was measured by CCK-8 assay, and the activation of protein kinase B (Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24, 48 and 72 h, and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells, the mRNA and protein levels of FAM3C were significantly increased in the WSU-HN6 cells (P<0.05). The viability of WSU-HN6 cells transfected with siFAM3C was significantly inhibited at 48 h and 72 h (P<0.05). siFAM3C treatment inhibited the activation of Akt (P<0.05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt (P<0.05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt (P<0.05). CONCLUSION: FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.     

Effect of beclin-1 gene silencing on TuAKe-injured gastric cancer SGC-7901 cells

AIM: To observe the effect of beclin-1 silencing by the technique of RNA interference on the injury of human gastric cancer SGC-7901 cell by Sheliugu extract (the extract from tuber of Amorphophallus konjac, TuAKe). METHODS: To knock down the expression of beclin-1 gene, SGC-7901 cells were transfected with lentiviral vector carrying beclin-1-shRNA. The beclin-1 gene knock-down and non-knock-down SGC-7901 cells were treated with TuAKe. The cell viability was analyzed by CKK-8 assay. The percentages of apoptotic cells were detected by flow cytometry. The expression of beclin-1 and LC3 was detected by Western blot. RESULTS: The beclin-1 gene silencing decreased the protein expression of beclin-1 and increased the protein expression of LC3 in the SGC-7901 cells, leading to the decrease in cell viability and the increase in apoptotic rate (P<0.05). TuAKe increased the protein expression of beclin-1 and LC3 in the SGC-7901 cells, and decreased the protein expression of LC3 in the SGC-7901 cells with beclin-1 gene silencing, thus inhibiting the cell viability and increasing the apoptotic rate (P<0.05). CONCLUSION: Beclin-1 gene silencing inhibits the activation of beclin-1-related signaling pathway in gastric cancer SGC-7901 cells, and aggravates the injury of cell viability induced by TuAKe.     

PPARγ mediates effects of diosgenin on proliferation and apoptosis in human glioblastoma U87MG cells

AIM:To investigate the effect of diosgenin (Dio) on the proliferation, apoptosis and expression of peroxisome proliferator-activated receptor γ (PPARγ) in human glioblastoma U87MG cells and its possible mechanism. METHODS:Human astrocytes (HA) and U87MG cells were cultured in vitro and treated with Dio (0, 10, 20, 30, 40 and 50 μmol/L) and GW9662 (5 μmol/L) for 48 h, and then the cell viability was detected by CCK-8 assay. Cell colony formation assay was used to assess the proliferation potential. Flow cytometry was used to analyze the cell cycle distribution and apoptosis. The mRNA expression level of PPARγ was measured by RT-PCR. Western blot was used to determine the protein levels of PPARγ, cyclin D1, cyclin E1, Bcl-2 and Bax. RESULTS:Dio had no significant influence on the viabi-lity of HA (P>0.05). However, Dio remarkably reduced the viability of U87MG cells in a dose-dependent manner (P<0.05) with IC₅₀ of 24.31 μmol/L. Meanwhile, Dio remarkably diminished colony formation ability (P<0.05), induced G₀/G₁ phase arrest of the cell cycle and apoptosis (P<0.05), up-regulated the expression of PPARγ at mRNA and protein levels, increased the protein level of Bax (P<0.05), and down-regulated the protein levels of cyclin D1, cyclin E1 and Bcl-2 (P<0.05) in a dose-dependent manner. However, these effects induced by Dio were inhibited by GW9662 (P<0.05), a specific inhibitor of PPARγ. CONCLUSION:Dio may inhibit proliferation and induce apoptosis in human glioblastoma U87MG cells most likely via up-regulating the expression of PPARγ, and then down-regulating the protein levels of cyclin D1, cyclin E1 and Bcl-2, and up-regulating the protein level of Bax.     

miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.     

27-Hydroxycholesterol modulates lung cancer cell proliferation by activation of LXR signaling pathway

AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway.     

蛋白质激发子Hrip1对小麦黄矮病的诱抗作用

小麦黄矮病是由大麦黄矮病毒（barley yellow dwarf virus，BYDV）引起的一种小麦病毒病，其传播介体是小麦蚜虫，在小麦生产中造成巨大的经济损失。近年来，植物诱导抗性作为一种新兴的植物病虫害防治措施引起了广泛的关注。蛋白质激发子Hrip1可以激活多种植物的免疫防御反应诱导植物产生广谱抗性。本研究评价了Hrip1对小麦黄矮病的诱抗效果。用30 μg/mL的Hrip1溶液进行小麦浸种和幼苗喷雾，随后接种BYDV，接种后第14 d，Hrip1对小麦黄矮病控制效果在50%以上，接种后第21 d控制效果仍在30%以上。实时荧光定量PCR检测结果显示，在Hrip1处理的小麦幼苗体内，BYDV外壳蛋白mRNA的数量显著低于对照组；EPG结果显示，在Hrip1处理的小麦幼苗上，麦二叉蚜寻找叶片刺吸位点和韧皮部取食位点的时间增加。以上结果表明：Hrip1能够有效地抑制BYDV在小麦体内的增殖；影响传毒媒介麦二叉蚜的取食行为，抑制其传毒能力。此外，Hrip1处理小麦能有效缓解BYDV引起的叶片黄化和植株矮化的症状。因此，Hrip1可以作为生物诱导剂综合控制小麦黄矮病。     

表观遗传修饰在骨骼肌细胞增殖分化过程中的研究进展

骨骼肌是肌肉的主要构成部分，骨骼肌细胞发生增殖和分化的过程都是肌肉发育的基础，直接影响着家养动物的产肉性能。研究发现表观遗传修饰作用对骨骼肌细胞增殖分化具有重要的调控作用，表明该遗传修饰作用对家养动物肌肉发育具有重大的意义。作者从DNA甲基化对骨骼肌细胞增殖分化影响、组蛋白乙酰化所含因子调控基因选择表达作用、非编码RNA调控和染色体重塑作用所起的影响等方面分别介绍了表观遗传在骨骼肌细胞增殖分化过程中的研究进展，简述了不同修饰方式和不同作用因子对骨骼肌增殖和分化两个过程的影响。同时也回顾了前人在研究骨骼肌增殖分化过程所用到的方法和手段，进而分析了表观调控作用因子在骨骼肌生长过程中所起到的作用。旨在进一步阐述表观遗传修饰在骨骼肌增殖和分化过程中所起到的重要作用，增强对骨骼肌增殖分化调控过程的了解，为和动物生产实际相结合提供参考途径，同时也为骨骼肌生长发育等分子调控提供更多参考素材。     

SNAT2介导蛋氨酸对牛乳腺上皮细胞增殖与自噬的调节作用

钠离子依赖性中性氨基酸转运体2（SNAT2）是一种氨基酸转运蛋白，可转运中性氨基酸，广泛分布于多种细胞中。氨基酸既可作为蛋白质合成的底物，也是调节细胞新陈代谢的关键信号分子，但SNAT2是否介导氨基酸调节BMECs增殖和自噬尚未见报道。本研究利用CASY细胞计数和Western blotting技术检测SNAT2过表达和siRNA干扰后牛乳腺上皮细胞（BMECs）增殖情况以及SNAT2对自噬标志蛋白LC3-Ⅰ/Ⅱ表达量的影响，并利用免疫荧光检测细胞自噬斑点（LC3-Ⅱ）变化。结果显示，SNAT2过表达时，p-PI3K、p-mTOR和Cyclin D1表达量增加，反之，p-PI3K、p-mTOR和Cyclin D1表达量下降。SNAT2抑制时，LC3-Ⅱ表达量增加，免疫荧光检测自噬斑点增多。添加自噬增强剂海藻糖（trehalose，Tre）和蛋氨酸（methionine，Met）后，与单一添加Tre组相比，Met+Tre组p-mTOR表达量增加，LC3-Ⅱ表达量降低，胞浆内绿色自噬斑点减少；添加Tre和Met并抑制SNAT2时，p-mTOR表达量下降，LC3-Ⅱ表达量增多，胞浆内绿色自噬斑点增加。以上结果表明，SNAT2可介导Met通过调控PI3K-mTOR/Cyclin D1信号通路调节BMECs的增殖与自噬。     

F-box domain is involved in regulating the proliferation of MCF-7 cells by FBXO39 protein

AIM: To investigate the effect of F-box domain on the regulation of MCF-7 cell proliferation by FBXO39 protein. METHODS: The effect of F-box domain on the localization of FBXO39 protein in the MCF-7 cells was investigated. MCF-7 cell cDNA library was used as the template resource. The full-length cDNA sequence of FBXO39 was amplified by PCR method and subcloned into eukaryotic expression vector pEGFP-C2. The pEGFP-FBXO39ΔF (F-box domain deletion mutation) plasmid was successfully constructed with the template resource of pEGFP-FBXO39 plasmid. The recombinant plasmids were transfected into the MCF-7 cells, and then the expression of FBXO39 and FBXO39ΔF were determined by Western blot. The cellular localization of FBXO39 and FBXO39ΔF were observed by confocal microscopy. The localization of endogenous FBXO39 in the MCF-7 cells was detected by immunofluorescence staining. In addition, MTT and EdU assays were used to measure the cell proliferation, flow cytometry was used to measure the cell cycle distribution, and immunohistochemical staining was used to observe the expression of FBXO39 in the breast cancer and para-carcinoma tissues. RESULTS: The eukaryotic expression vector pEGFP-FBXO39 and pEGFP-FBXO39ΔF were constructed successfully. F-box domain had no effect on the cell localization of FBXO39. FBXO39 promoted MCF-7 cell proliferation but FBXO39ΔF did not. FBXO39 was highly expressed in the breast cancer tissues. CONCLUSION: F-box domain had no effect on the cellular localization of FBXO39 protein. However, it plays an important role in the biological function of FBXO39. FBXO39 may be related to breast cancer tumorigenesis.