RNA干扰下调p53抑制因子iASPP表达对膀胱癌细胞的影响

Objective To discuss the effects of silencing of iASPP gene on human bladder cancer cells. Methods RNAi silencing of iASPP gene in bladder cancer cell 5637 and T24 cells were used by lentiviral mediated interfering short hairpin RNAs. Cell proliferation was tested by MTT assay, and rate of colony was tested by colony formation assay. Cell cycles were tested by using fluorescence-activated cell sorting. Results Down-regulation of iASPP could inhibit the growth and proliferation of human bladder cancer cells (P＜0.05). iASPP know-down could decrease the colony formation of 5637 and T24 cells (P＜0, 05). Knocking down of iASPP in 5637 and T24 cells showed cell arrested at G1. Conclusions Silencing of iASPP gene could inhibit proliferation and colony formation of bladder cancer, iASPP might be an important target for gene therapy of bladder cancer.     

RNA干扰下调p53抑制因子iASPP表达对膀胱癌细胞的影响

目的 探讨RNA干扰使p53抑制因子iASPP表达下调对膀胱癌细胞的影响.方法 iASPP siRNA慢病毒感染入膀胱癌细胞株5637和T24,实时定量PCR和蛋白质印迹法检测iASPP的表达;噻唑盐法测定细胞生长;集落形成测定法检测集落形成比率;荧光激活细胞分选术检测细胞周期.结果 iASPP siRNA慢病毒感染后细胞iASPP mRNA为0.60±0.02,低于阴性对照慢病毒组(1.00±0.03,P＜0.05).膀胱癌细胞iASPP蛋白表达降低.iASPP下调能抑制入膀胱癌细胞5637和T24的生长和增殖(P＜0.05),每个集落中细胞数显著减少(P＜0.05),集落数目也低于阴性对照组(P＜0.05).iASPP siRNA慢病毒5637细胞培养中的G1期细胞比率明显高于阴性对照慢病毒组,分别为(51.94±0.98)%和(46.00±0.77)%;T24细胞结果与之类似,分别为(60.04±0.45)%和(53.62±0.69)%;组间比较差异均有统计学意义(P＜0.05).结论 RNA干扰能够使膀胱癌细胞iASPP表达下调,从而抑制膀胱癌细胞的生长和增殖.

Abstract：

Objective To discuss the effects of silencing of iASPP gene on human bladder cancer cells. Methods RNAi silencing of iASPP gene in bladder cancer cell 5637 and T24 cells were used by lentiviral mediated interfering short hairpin RNAs. Cell proliferation was tested by MTT assay, and rate of colony was tested by colony formation assay. Cell cycles were tested by using fluorescence-activated cell sorting. Results Down-regulation of iASPP could inhibit the growth and proliferation of human bladder cancer cells (P＜0.05). iASPP know-down could decrease the colony formation of 5637 and T24 cells (P＜0, 05). Knocking down of iASPP in 5637 and T24 cells showed cell arrested at G1. Conclusions Silencing of iASPP gene could inhibit proliferation and colony formation of bladder cancer, iASPP might be an important target for gene therapy of bladder cancer.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cell Colo205 in vitro

Objective To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against β-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205. Methods The shRNA plasmid vectors against β-catenin were constructed and transfected into Colo205 ceils with LipofectamineTM 2000. The expression of β-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the β-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay. Results The shRNA vectors targeted against β-catenin were successfully constructed and efficiently suppressed the expression of β-catenin mRNA and protein (P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of β-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different (P<0.05). Conclusions The specific shRNAs targeted against β-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.     

Faculty of Anaesthetists of the Royal College of Surgeons of Ireland

The Faculty of Anaesthetists of the Royal College of Surgeons of England, 35–43 Lincoln's Inn Fields, London WC2A 3PN. Telephone: 01-405 3474.     

深圳市某两所小学流行性腮腺炎突发疫情的流行病学分析

目的：通过对深圳市某两所小学发生的流行性腮腺炎突发疫情的流行病学特点及差异性进行分析,为制定科学、高效的防控策略提供科学依据。方法2013年5～7月深圳市大鹏新区某两所小学爆发流行性腮腺炎,以学校为整体研究对象,分别标记为学校A（24个班,学生1210例）和学校B（27个班,学生1274例）,对比两所小学的疫情流行病学差异性。结果分析发现,学校A流行性腮腺炎发病率为4.30%,发病班级所占比54.17%,均较学校B1.73%和29.63%高,对比差异有统计学意义（P＜0.05）;分析显示学校A学生出现疫病平均年龄为（11.2±1.1）岁,较学校B（9.34±1.0）岁,对比差异明显（P＜0.05）;且两组疫病患儿在接种疫苗率对比上差异无统计学意义（P＞0.05）;但疫情发生时,学校B疫苗紧急接种率明显高于学校A,对比差异有统计学意义（P＜0.05）。结论小学作为流行性腮腺炎爆发的主要场所之一,疫病爆发高峰季节前,针对易感染人群给予相应的疫苗接种等预防控制措施,同时加强流行性腮腺炎的监测,对于降低感染人群数量,减轻、遏制疫情有着积极的意义,值得相关防控部门重视。