Triad1调节K562细胞增殖、凋亡和耐药研究*

吴蕾蕾**,徐亚,刘

(南通大学附属医院血液内科,江苏226001

[] 目的:研究Triad1对慢性髓细胞白血病(CML)细胞株K562增殖和凋亡的作用以及与伊马替尼(IM)耐药的关系。方法:将K562细胞进行血清饥饿培养,流式细胞仪检测细胞周期变化,蛋白免疫印迹法检测PCNA等增殖相关蛋白的表达。shRNA干扰K562细胞后细胞周期和凋亡的变化。构建耐药细胞K562R,蛋白免疫印迹法检测K562K562R细胞中Triad1以及凋亡相关蛋白的表达。结果:(1)血清饥饿培养后K562细胞阻滞在G0/G1期,恢复血清培养后随着K562细胞增殖,Triad1表达减少,而PCNA等蛋白表达增加。(2)干扰Triad1表达后,K562细胞增殖能力增强,G1期细胞减少。(3K562R细胞Triad1的表达下降,凋亡相关蛋白表达降低。结论:Triad1通过调节细胞周期来调节K562细胞的增殖,干扰Triad1可促进K562细胞增殖,凋亡减少,Triad1表达下降可能与K562细胞耐药相关。

[关键词] 慢性髓细胞白血病;K562细胞;Triad1;增殖;凋亡;耐药

[中图分类号] R733.7 [文献标志码] A

Triad1 regulates cell proliferation, apoptosis, and chemo-resistance In K562 cells

WU Leilei, XU Ya, LIU Hong

(Department of Hematology, Affiliated Hospital of Nantong University, Jiangsu 226001)

[Abstract] Objective: o investigate the effects of Triad1 on the proliferation and apoptosis of K562 cells in chronic myeloid leukemia (CML) and its relationship with imatinib (IM) -mediated CML cell resistance, and to explore its mechanism and significance. Methods: The K562 cells underwent serum starvation. Cell cycle progression was determined by flow cytometry. The changes in expression of PCNA in the cell proliferation was measured by Western blot analysis. K562 cells were transfected with either triad1 shRNA or control shRNA. Triad1 level was significantly decreased with shRNA transfection, which was determined by Western blot. Flow cytometry was used to measure cell cycle distribution and apoptosis. K562 cells were added into imatinib (IM) according to different concentration gradients to construct imatinib (IM) resistant cell K562R. Western blot analysis was used to detect the level of Triad1 and related protein. Results(1)After serum deprivation in K562 cells, the cell cycle was arrested in G0 / G1 phase after re-feeding. With the stimulation of serum in K562 cells, the expression of Triad1 was decreased. The expression of PCNA is increased.(2)Cell proliferation ability was promoted when expression of Triad1 was lost. Cells in G1 phase were decreased. (3)The expression of Triad1 was decreased with imatinib resistance in K562R, and the level of apoptosis-related protein was decreased. Conclusions: Triad1 regulated the proliferation of K562 cells via the regulation of cell cycle. Knockdown of Triad1 promoted the proliferation of K562 cells. Triad1 may promote apoptosis in K562 cells. The expression of Triad1 is related to the resistance of K562 cells.

[Key words] chronic myeloid leukemia; K562 cells; Triad1; proliferation; apoptosis; resistance

Wnt3蛋白在非小细胞肺癌中的表达及临床意义

董业峰1*,马剑波2

1海安县人民医院检验科,江苏226600;2南通大学附属医院肿瘤放疗科)

[摘 要] 目的:研究Wnt3蛋白在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达水平及临床意义。方法:采用免疫组织化学方法对64例NSCLC患者术后标本进行Wnt3蛋白表达水平的检测。结合阳性细胞染色强弱及百分比计算Wnt3免疫组化染色评分,分析Wnt3表达与患者临床病理特征之间的关系,并探讨Wnt3蛋白在NSCLC患者中表达的意义。结果:在NSCLC组织中,Wnt3的免疫组化染色定位于胞浆中,其阳性表达率为64.1%(41/64),显著高于癌旁组织阳性表达率40.0%(12/30)(P=0.028)。Wnt3蛋白表达强度与NSCLC的分化程度(P=0.028)、淋巴结转移(P=0.008)和病理分期(P=0.000)有显著相关性,而与NSCLC的年龄、性别、吸烟状态、组织类型及侵润深度(T分期)无关(P均>0.05)。结论:Wnt3蛋白在NSCLC中高表达,可能与NSCLC的发生发展相关。

[关键词] 非小细胞肺癌;Wnt3蛋白;免疫组织化学

[中图分类号] R734.2 [文献标志码] A

The expression and clinical significance of Wnt3 in non-small cell lung cancer

DONG Yefeng1, MA Jianbo2

(1Department of Laboratory Medicine, Hai’an County People’s Hospital, Jiangsu 226600;

2Department of Oncology Radiotherapy, the Affiliated Hospital of Nantong University)

[Abstract] Objective: To study the expression and clinical significance of Wnt3 in non-small cell lung cancer (NSCLC). Methods: Immunohistochemistry was used to detect the expression of Wnt3 protein in 64 specimens of NSCLC patients. The expression of Wnt3 immunohistochemical staining was calculated with the percentage and staining intensity index of positive cells staining. Chi-square test was used to analyze the relationship between Wnt3 expression and clinicopathological features of the patients, and the correlation of Wnt3 expression in NSCLC patients was explored. Results: In NSCLC tissues, the immunohistochemical staining of Wnt3 was localized in the cytoplasm, and the positive expression rate of Wnt3 was 64.1% (41/64), significantly higher than that of paracancerous tissues,which was 40.0% (12/30) (P=0.028). The expression of Wnt3 protein was significantly correlated with the degree of NSCLC differentiation (P=0.028), lymph node metastasis (P=0.008) and pathological stage (P=0.000).However,there was no significant association with age, gender, smoking status, tissue type, and NSCLC infiltration depth. Conclusion: Wnt3 protein is highly expressed in NSCLC and may be related to the development of NSCLC.

[Key words] non-small cell lung cancer; Wnt3; immunohistochemistry

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低浓度氯气诱导大鼠喉部损伤及地塞米松治疗效果*

**,吴

(常熟市第二人民医院耳鼻咽喉科,江苏 215500

[] 目的:观察低浓度氯气吸入后大鼠喉部损伤及地塞米松的治疗效果。方法:大鼠置于浓度为50 ppm氯气容器罐中15 min,喉镜观察大鼠喉部的损伤,病理检查喉部组织学改变,评估即刻地塞米松治疗后的效果。结果:大鼠吸入低浓度氯气后喉部出现炎性水肿渗出损伤改变,即刻地塞米松治疗后,喉部粘膜水肿消退明显,分泌物明显减少,病理示被覆鳞状上皮完整,细胞形态大小正常,表面未见有炎性渗出,无出血,粘膜下固有层未见明显水肿,软骨形态正常,未见粘液变性。结论:早期给予地塞米松治疗可有效缓解较低浓度氯气吸入后导致的喉部损伤。

[关键词] 喉损伤;低浓度氯气;地塞米松;大鼠

[中图分类号] R767.8 [文献标志码] A

The low concentration of chlorine induced laryngeal injury and dexamethasone treatment

LU Jia, WU Zhen

(Department of Otolaryngology, the Second People’s Hospital of Changshu, Jiangsu 215500)

[Abstract] Objective: To investigate the inflammatory changes of laryngeal injury in rats after inhaling the low concentration of chlorine. To explore the therapeutic effect of laryngeal injury by dexamethasone. Methods:To observe the fibrolaryngoscope and the pathological changes of larynx in rats after 15 minutes in the chlorine tank with the concentration of 50PPM. The changes of laryngeal were observed after withdrawal of chlorine treated by dexamethasone (100mg/kg) immediately. Results:The low concentration of chlorine inhalation could induce the inflammatory edema and exudation of larynx in rats. After being treated by dexamethasone, Laryngeal mucosa edema subsided significantly, and secretion decreased significantly. Covered squamous epithelium was integrated, The shape and the size of cells were normal.There was no inflammatory exudate on the surface, or no bleeding. There was not any submucosal lamina propria edema.Cartilage morphology was normal, and there was not mucous degeneration. Conclusion:Dexamethasone can effectively alleviate the laryngeal damage caused by low concentration of chlorine at the early stage.

[Key words] chlorine; larynx injury; dexamethasone; rat