Sugarcane (Saccharum spp.) is an important sugar crop. Biotic and abiotic stresses such as diseases, cold, and drought are major factors limiting sugarcane production. β-1,3-glucanase (EC 3.2.1.39), a member of the pathogenesis-related protein family, plays an essential role not only in the plant defenses against pathogens but also in plant growth, development, and abiotic stress responses. To systematically investigate the sugarcane β-1,3-glucanase gene family, 132 glycoside hydrolase (GH) 17 family members were identified in the genomes of the sugarcane wild species Saccharum spontaneum 'Np-X', the tropical species S. officinarum 'LA-Purple', and the Saccharum spp. hybrid cultivar 'R570'. The results of the phylogenetic analysis categorized them into four subfamilies, of which subfamily Ⅳ had the largest proportion of members (102). The members of the sugarcane GH17 gene family contained five conserved motifs and 0-16 introns. The majority of the GH17 genes exhibited a genome-wide replication pattern, with 89.50% originating from S. spontaneum 'Np-X' and S. officinarum 'LA-Purple', while 58.10% of them in the Saccharum spp. hybrid cultivar 'R570' belonged to the discrete replication type. Four major classes of cis-acting elements were identified in the promoters, including the elements related to plant growth, development, and tissue-specific expression (14.21%), light-responsive elements (38.24%), biotic or abiotic stress-responsive elements (9.18%), and hormone-responsive elements (38.37%), which suggested that this gene family was involved in plant growth, development, hormone responses, and stress responses. Transcriptome and quantitative real-time PCR (RT-qPCR) analyses showed that the sugarcane GH17 genes exhibited tissue-specific expression and were differentially expressed under low temperature, drought, and hormone treatments, as well as during the interactions between different sugarcane genotypes and Sporisorium scitamineum, suggesting their potential roles in plant defenses. In addition, some SsGlu genes (SsGlu5, SsGlu20, SsGlu21, SsGlu25, SsGlu28, and SsGlu39) were expected to serve as candidate stress-related genes. This study lays a foundation for further revealing the molecular mechanisms of the stress resistance of sugarcane via β-1,3-glucanase genes.
Saccharum/physiology* ; Stress, Physiological/genetics* ; Glucan 1,3-beta-Glucosidase/metabolism* ; Multigene Family ; Phylogeny ; Gene Expression Regulation, Plant ; Plant Proteins/genetics*

Objective To investigate the effects of interleukin-17 (IL-17) on the cell proliferation, apoptosis and migration of human laryngeal carcinoma Hep-2 cells.Methods IL-17 was transiently transfected into Hep-2 cells, and at the same time empty vector group (pEGFP-N1) and normal control group were set up.The efficiency of transfection was evaluated by fluorescence microscope, and the mRNA and protein expressions of IL-17 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.The proliferation of cells was detected by methyl thiazolyl tetrazolium (MTT) method, and the apoptosis was detected by flow cytometry.The migration ability was detected by wound-healing assay and Transwell assay.ResultsHep-2 cells transfected with empty vector pEGFP-N1 and IL-17 showed green fluorescence under the fluorescence microscope.Hep-2 cells expressed IL-17 at both mRNA and protein levels after transfection with IL-17.Compared with the normal control group, the proliferation of IL-17 transfected Hep-2 cells was significantly inhibited after 48 h transfection (0.34±0.03 vs.0.46±0.04, P=0.006).The apoptotic rate of IL-17 transfected cells was higher than that of normal control group (26.80%±0.80% vs.2.90%±0.31%, P=0.000).According to the wound-healing assay, compared with the normal control group, the scratch width of IL-17 transfected cells was significantly greater (1.59±0.01 vs.1.36±0.01, P=0.000).Transwell migration experiment showed that the migration of IL-17 transfected cells was significantly lower than that of the normal control group (26.33±2.08 vs.49.33±1.53, P=0.000).Conclusion IL-17 can inhibit the proliferation of human laryngeal carcinoma Hep-2 cells, reduce their migration ability and enhance their apoptosis ability.Therefore, IL-17 may inhibit the occurrence and development of laryngeal carcinoma through a variety of mechanisms.

OBJECTIVE: To investigate the mRNA and protein expression of MICA in laryngeal squamous cell carcinoma tissue and the Hep-2 cells. METHOD: Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blot were used to detect the expression of MICA mRNA and protein levels in the Hep-2 cells and laryngeal cancer tissues. RESULT: The MICA mRNA showed higher expression in Hep-2 cells by RT-PCR. Compared with the control, the mRNA expression of MICA was significantly enhanced in laryngeal cancer tissues (t = 11.878, P < 0.01). The intensity of MICA expression is not related to the clinical stage of cancer. MICA protein demonstrated higher level expression by Western blot. The intensity of MICA protein expression was decreased with increased clinical stage in laryngeal cancer tissues. CONCLUSION The MICA mRNA showed stronger expression in Hep-2 cells and laryngeal cancer tissues. The intensity of its expression is not related to clinical stage of cancer. The MICA protein expression was strong in Hep-2 cells. The intensity of MICA protein expression was decreased with increased clinical stage in laryngeal cancer tissues. MICA may play an important role in laryngeal carcinoma process.
Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Head and Neck Neoplasms ; metabolism ; pathology ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Squamous Cell Carcinoma of Head and Neck

Objective To investigate the effect of miR-27a mimic and inhibitor on proliferation and apoptosis in melanoma cell line WM239. Methods The miR-27a mimic,inhibitor and its negative control were transfected into WM239 cells. The transfection efficiency was evaluated by fluorescence microscope. The expres-sion of miR-27a was detected by real-time fluorescent quantitative PCR. The proliferation of cells was detected by MTT. The cell apoptosis and cell cycle were analyzed by flow cytometry. Results The transfection efficiency in WM239 cells was 80% to 90%. The expression of miR-27a was markedly up-regulated in miR-27a mimic group (2-△△CT value is 26. 98 ± 0. 01),with statistically significant difference(t= -1 123. 67,P=0. 00);and the miR-27a inhibitor group showed lower expression of miR-27a(2-△△CT value is 0. 96 ± 0. 02),there was no statisti-cally significant difference compared with normal control group(t=0. 04,P=0. 06). The proliferation of cells was obviously inhibited in miR-27a mimic group,and there was statistically significant difference compared with normal control group[absorbance of 72 h(0. 45 ± 0. 02)∶(0. 72 ± 0. 01),F=129. 56,P﹤0. 05]. The percent-age of WM239 cells in G0-G1 phase was increased[(74. 83 ± 1. 46)∶(63. 73 ± 1. 25),F=30. 33,P﹤0. 05], and the percentage of WM239 cells in S phase and G2-M phase were decreased[(21. 33 ± 1. 75)∶(27. 50 ± 1. 25),F=14. 98,P﹤0. 05;(3. 90 ± 1. 31)∶(8. 80 ± 2. 10),F=3. 66,P﹤0. 05]. The apoptosis rate of cells was significantly increased in miR-27a mimic group compared with normal group[(29. 67 ± 0. 91)%∶(1. 44 ± 0. 85)%,F=530. 90,P﹤0. 01],but the inhibitor group had no obvious effect on cell cycle and cell apoptosis. Conclusion MiR-27a can suppress melanoma cell proliferation and act as a tumor suppressor gene,which is rel-evant to induce cell apoptosis and block cell cycle in G0-G1 phase.

Objective To investigate the effects of resveratrol (Res) on the proliferation,cell cycle and apoptosis of three breast cell lines DU4475,MDA-MB-23land MCF-7 with different estrogen receptor (ER) expressions.Methods Res was added to the drug treatment group at 6.25,12.5,25,50,100 and 200 μmol/L,respectively.Three observation periods at 24,48 and 72 hours were set up respectively.MTT assay was used to detect the effects of Res on proliferation of breast cells.Cell cycle and apoptosis were analyzed by flow cytometry (FCM).The concentrations of drugs were 0,12.5,25,50 μmol/L.The results were analysed by the statistical software SPSS17.0.Results After Res intervention,the proliferations of three cell lines were inhibited to different extent.After 48 and 72 hours of Res,inhibitions of Res with different concentration were significant different (F =15.26,P ＜ 0.05).Inhibition rates of DU4475 and MDA-MB-231 with ER-negative were higer than that of MCF-7 with ER-positive.FCM results prompted that these three kinds of cells were blocked in S phase after 48 hours treatment of 12.5-50 μmol/L Res and the block percentages had significant difference (F =34.81,P ＜ 0.05).The percentages of S phase for DU4475 and MDA-MB-231 arresting were higher than that of MCF-7.For DU4475,the apoptotic and necrosis percentage were higher than that of MCF-7 at 100 μmol/L (t =16.082,P ＜0.05).Conclusion Res can inhibit proliferation,induce cell cycle changing and apoptosis of DU4475,MDA-MB-231 and MCF-7 cells.The inhibitions of Res on DU4475 and MDA-MB-231 are better than that of MCF-7 with ER-positive.