Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

Hyperoxia-induced apoptosis of AEC Ⅱ in premature rats by inhibiting expression of focal adhesion kinase

Objective To explore the role of focal adhesion kinase (FAK) in hyperoxia-apopto- sis of type Ⅱ alveolar epithelial cells (AEC Ⅱ s) of premature rats. Methods AEC Ⅱ from prema- ture rat lungs were cultured and randomly assigned to air group and hyperoxia group. After exposed to hyperoxia for 6, 12, 24 and 48 h, apoptosis rate of AEC Ⅱ were analyzed by flow cytometry with an- nexin-Ⅴ/propidium iodine (PI) double staining. FAK mRNA and FAK and fAK-Tyr397 peptide were detected by RT-PCR and Western blot, respectively. Results Positive cells of Annexin-Ⅴ+ / PI- in AEC Ⅱ after 6,12,24 and 48 h of hyperoxia exposure were significantly decreased, and the maximal apoptosis rate of AEC Ⅱ (stained of Annexin-Ⅴ+ / PI- ) was found in the hyperoxia group at 12 h (23.83%±4.43%). Compared to the air group, the expression of FAK mRNA and of FAK de- creased markedly and progressively in hyperoxia groups at 12, 24 and 48 h(P<0. 05). Conclusions Decreased expression of FAK induced by hyperoxia is likely to contribute to the apoptosis and neco-z sis of AEC Ⅱ.     

高浓度氧通过抑制黏着斑激酶表达损伤早产大鼠肺泡Ⅱ型细胞

目的 探讨高浓度氧暴露不同时间点早产鼠肺泡Ⅱ型上皮细胞(AECⅡ)凋亡规律及其与黏着斑激酶(FAK)表达的关系.方法 原代培养早产鼠AECⅡ,暴露于高氧环境中6、12、24和48 h,并以空气组作为对照组,采用Annexin-Ⅴ和PI双标法经流式细胞仪检测AECⅡ凋亡情况,并采用Western印迹、RT-PCR技术分析AECⅡFAK多肽、磷酸化FAK(FAK-Tyr397)多肽和FAKmRNA表达变化.结果 与空气组比较,高氧暴露12 h,Annexin-Ⅴ+/PI(早期凋亡)亚群细胞所占比例最高,达(23.83±4.43)%.随高氧暴露时间延长,Annexin-Ⅴ+/PI-亚群细胞所占比例逐步减低,而Annexin-Ⅴ+/PI+亚群细胞所占比例逐步增高(P<0.05或P<0.01).AEC Ⅱ FAK、FAK-Tyr397多肽及FAK mRNA表达水平随高氧暴露时间延长呈明显下降趋势(P<0.05或P<0.01).结论 高浓度氧抑制FAK表达可能是AECⅡ凋亡和坏死的重要原因之一.     

Expression of glyoxalase Ⅰ and its effect on cell proliferation and apoptosis in endometrial carcinoma

Objective To examine the expressions of glyoxalase Ⅰ (GLO-Ⅰ ) in endometrial cancer tissues and cell lines and to investigate the roles of GLO-Ⅰ on proliferation and apoptosis in endometrial cancer cells. Methods Immunohistochemistry, western blot and RT-PCR were used to investigate the expressions of GLO-Ⅰ protein and mRNA in endometrial cancer tissues and Ishikawa cell lines ;enzyme activity of GLO-Ⅰ in normal endometrium, endometrial cancer and paraneoplastic tissue samples was detected with spectrophotometer; proliferation and apoptosis of Ishikawa cell before and after RNA interference (RNAi) procedure were detected by the methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. Results (1)There were significant differences of GLO-Ⅰ expression between normal endometrium (0/19) and endometrial cancer tissues ( 76%, 22/29 ); these were also significant differences of enzyme activity of GLO-Ⅰ among normal endometrium, paraneoplastic and endometrial cancer tissues( 1.1,0.8 vs 92.3 IU/mg; P <0.01 ). Enzyme activity of GLO-Ⅰ in fresh normal endometrium and paraneoplastic tissues was weak, while that of fresh endometrial cancer tissues was as high as 92. 3 IU/mg in average. (2)The expression of GLO-Ⅰ mRNA in Ishikawa cell transfected with GLO-Ⅰ siRNA was significantly lower than that in negative group (0.25 ± 0.06 vs 0.93 ± 0.10, P < 0.0l ), and the similar results that in the expression of GLO-Ⅰ protein (0.38 ±0.06 vs 0.94 ±0.13, P <0.01 ). (3) Proliferation in Ishikawa cell was significantly inhibited after silencing RNA expression of GLO-Ⅰ ( P = 0.028 ). The apoptosis rate of cells transfected with GLO- Ⅰ siRNA was significantly higher than that of negative control group and blank control group [ ( 6.7 ± 0.8 ) % vs ( 1.2 ± 0.4) %, ( 1.4 ± 0.4 ) %; P < 0.01 ]. Conclusion The expression and enzyme activity of GLO- Ⅰ is significantly increased in endometrial cancer, which could promote abnormal proliferation and inhibit apoptosis in endometrial cancer cells.     

Expression of glyoxalase Ⅰ and its effect on cell proliferation and apoptosis in endometrial carcinoma

Objective To examine the expressions of glyoxalase Ⅰ (GLO-Ⅰ ) in endometrial cancer tissues and cell lines and to investigate the roles of GLO-Ⅰ on proliferation and apoptosis in endometrial cancer cells. Methods Immunohistochemistry, western blot and RT-PCR were used to investigate the expressions of GLO-Ⅰ protein and mRNA in endometrial cancer tissues and Ishikawa cell lines ;enzyme activity of GLO-Ⅰ in normal endometrium, endometrial cancer and paraneoplastic tissue samples was detected with spectrophotometer; proliferation and apoptosis of Ishikawa cell before and after RNA interference (RNAi) procedure were detected by the methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. Results (1)There were significant differences of GLO-Ⅰ expression between normal endometrium (0/19) and endometrial cancer tissues ( 76%, 22/29 ); these were also significant differences of enzyme activity of GLO-Ⅰ among normal endometrium, paraneoplastic and endometrial cancer tissues( 1.1,0.8 vs 92.3 IU/mg; P <0.01 ). Enzyme activity of GLO-Ⅰ in fresh normal endometrium and paraneoplastic tissues was weak, while that of fresh endometrial cancer tissues was as high as 92. 3 IU/mg in average. (2)The expression of GLO-Ⅰ mRNA in Ishikawa cell transfected with GLO-Ⅰ siRNA was significantly lower than that in negative group (0.25 ± 0.06 vs 0.93 ± 0.10, P < 0.0l ), and the similar results that in the expression of GLO-Ⅰ protein (0.38 ±0.06 vs 0.94 ±0.13, P <0.01 ). (3) Proliferation in Ishikawa cell was significantly inhibited after silencing RNA expression of GLO-Ⅰ ( P = 0.028 ). The apoptosis rate of cells transfected with GLO- Ⅰ siRNA was significantly higher than that of negative control group and blank control group [ ( 6.7 ± 0.8 ) % vs ( 1.2 ± 0.4) %, ( 1.4 ± 0.4 ) %; P < 0.01 ]. Conclusion The expression and enzyme activity of GLO- Ⅰ is significantly increased in endometrial cancer, which could promote abnormal proliferation and inhibit apoptosis in endometrial cancer cells.     

Notch基因细胞内区在子宫颈癌组织中的表达及DAPT对子宫颈癌细胞的影响

Objective To study the expression and clinical significance of Notch intracellular domain (NICD) in cervical cancer and the effects of N-[N-(3,5-difluorophenyl)acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis of cervical cancer cell lines. Methods Western blot was used to detect the expression of NICD in the tissues of 40 cervical cancers and 21 normal cervix and its relationship with clinical features of cervical cancer was also analyzed. Proliferation of SiHa and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in SiHa and HeLa cells incubated with DAPT was detected by western blot. Results The expression level of NICD in cervical cancers was significantly higher than that of normal cervical tissues (1.237±0.353 vs 0.938±0.105, P<0.05). The NICD expression was higher in cervical cancers with high grade,lymph node involvement and parametrial invasion than that with low-middle grade (1.496±0.540 vs 1.150±0.216), without lymph node involvement (1.419±0.532 vs 1.159±0.210) and no parametrial invasion (1.718±0.710 vs 1.183±0.258), respectively (all P<0.05). The expression of NICD in cervical adenocarcinoma was higher than that of squamous cell cancer (1.463±0.395 vs 1.162±0.187, P<0.05). After SiHa and HeLa cells were incubated with DAPT, NICD expression was significantly lower than that in control (P<0.05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells was depended on its concentrations and times. Conclusions NICD may play a key role in the occurrence and progress of cervical cancer. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells may be due to decreased the formation of NICD.     

Notch基因细胞内区在子宫颈癌组织中的表达及DAPT对子宫颈癌细胞的影响

Objective To study the expression and clinical significance of Notch intracellular domain (NICD) in cervical cancer and the effects of N-[N-(3,5-difluorophenyl)acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis of cervical cancer cell lines. Methods Western blot was used to detect the expression of NICD in the tissues of 40 cervical cancers and 21 normal cervix and its relationship with clinical features of cervical cancer was also analyzed. Proliferation of SiHa and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in SiHa and HeLa cells incubated with DAPT was detected by western blot. Results The expression level of NICD in cervical cancers was significantly higher than that of normal cervical tissues (1.237±0.353 vs 0.938±0.105, P<0.05). The NICD expression was higher in cervical cancers with high grade,lymph node involvement and parametrial invasion than that with low-middle grade (1.496±0.540 vs 1.150±0.216), without lymph node involvement (1.419±0.532 vs 1.159±0.210) and no parametrial invasion (1.718±0.710 vs 1.183±0.258), respectively (all P<0.05). The expression of NICD in cervical adenocarcinoma was higher than that of squamous cell cancer (1.463±0.395 vs 1.162±0.187, P<0.05). After SiHa and HeLa cells were incubated with DAPT, NICD expression was significantly lower than that in control (P<0.05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells was depended on its concentrations and times. Conclusions NICD may play a key role in the occurrence and progress of cervical cancer. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells may be due to decreased the formation of NICD.