CAJ | 학술논문

目的探讨靶向沉默ALKBH3基因表达对人前列腺癌LNCaP细胞裸鼠皮下移植瘤生长的影响及其机制.方法建立人前列腺癌LNCaP细胞裸鼠皮下移植瘤模型,瘤体局部注射重组质粒ALKBH3-小干扰RNA(siRNA)作为实验组,空质粒载体为对照组,生理盐水为空白组.绘制移植瘤生长曲线,采用原位缺口末端标记法(TUNEL)检测移植瘤细胞的凋亡,采用免疫组织化学法和Western blot法检测移植瘤组织中ALKBH3、增殖细胞核抗原(PCNA)及B细胞淋巴瘤/白血病-2(bcl-2)的表达.结果实验组移植瘤的生长速度低于对照组和空白组.TUNEL法检测结果显示空白组和对照组的积分吸光度值分别为460.2±203.5和588.5±241.3,均低于实验组的积分吸光度值(6558.2±1 567.1,P＜0.05).免疫组织化学结果检测显示实验组移植瘤组织ALKBH3蛋白的表达水平(0.26±0.03)低于空白组(0.43±0.07)和对照组(0.48 ±0.06,P＜0.05);实验组移植瘤组织PCNA蛋白的表达水平(0.16±0.02)低于空白组(0.28±0.05)和对照组(0.31±0.05,P＜0.05);实验组移植瘤组织bcl-2蛋白的表达水平(0.31 ±0.05)低于空白组(0.55±0.12)和对照组(0.49±0.11,P＜0.05).Western blot检测结果显示实验组移植瘤组织ALKBH3蛋白的相对表达水平(0.16±0.05)低于空白组(0.78±0.27)和对照组(0.85±0.31,P＜0.05);实验组移植瘤组织PCNA蛋白的相对表达水平(0.13 ±0.02)低于空白组(0.49±0.17)和对照组(0.45 ±0.13,P ＜0.05),实验组移植瘤组织bcl-2蛋白的相对表达水平(0.23 ±0.04)低于空白组(0.41±0.15)和对照组(0.44±0.19,P＜0.05).结论靶向沉默ALKBH3基因表达可以诱导细胞凋亡,抑制人前列腺癌LNCaP细胞裸鼠皮下移植瘤的生长,其作用机制可能与抑制PCNA和bcl-2表达有关.
목적 탐토파향침묵ALKBH3기인표체대인전렬선암LNCaP세포라서피하이식류생장적영향급기궤제.방법 건립인전렬선암LNCaP세포라서피하이식류모형,류체국부주사중조질립ALKBH3-소간우RNA(siRNA)작위실험조,공질립재체위대조조,생리염수위공백조.회제이식류생장곡선,채용원위결구말단표기법(TUNEL)검측이식류세포적조망,채용면역조직화학법화Western blot법검측이식류조직중ALKBH3、증식세포핵항원(PCNA)급B세포림파류/백혈병-2(bcl-2)적표체.결과 실험조이식류적생장속도저우대조조화공백조.TUNEL법검측결과현시공백조화대조조적적분흡광도치분별위460.2±203.5화588.5±241.3,균저우실험조적적분흡광도치(6558.2±1 567.1,P＜0.05).면역조직화학결과검측현시실험조이식류조직ALKBH3단백적표체수평(0.26±0.03)저우공백조(0.43±0.07)화대조조(0.48 ±0.06,P＜0.05);실험조이식류조직PCNA단백적표체수평(0.16±0.02)저우공백조(0.28±0.05)화대조조(0.31±0.05,P＜0.05);실험조이식류조직bcl-2단백적표체수평(0.31 ±0.05)저우공백조(0.55±0.12)화대조조(0.49±0.11,P＜0.05).Western blot검측결과현시실험조이식류조직ALKBH3단백적상대표체수평(0.16±0.05)저우공백조(0.78±0.27)화대조조(0.85±0.31,P＜0.05);실험조이식류조직PCNA단백적상대표체수평(0.13 ±0.02)저우공백조(0.49±0.17)화대조조(0.45 ±0.13,P ＜0.05),실험조이식류조직bcl-2단백적상대표체수평(0.23 ±0.04)저우공백조(0.41±0.15)화대조조(0.44±0.19,P＜0.05).결론 파향침묵ALKBH3기인표체가이유도세포조망,억제인전렬선암LNCaP세포라서피하이식류적생장,기작용궤제가능여억제PCNA화bcl-2표체유관.
Objective To investigate the antitumor effect of small interfering RNA (siRNA)-mediated inhibition of ALKBH3 gene expression on nude mice xenograft with human prostate cancer cell,and to explore its mechanism of action.Methods The nude mice model of human prostate cancer was established by injecting human LNCaP cells subcutaneously.The ALKBH3-siRNA was injected into the tumor of experiment group.Terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) was undertaken to detect the cell apoptosis in the tumor tissue.Immunohistochemistry (IHC) and Western blotting was used to detect the expression of ALKBH3,proliferating cell nuclear antigen (PCNA) and B cell lymphoma/leukemia-2 (bcl-2).Results The tumor growth of experiment group was slower than that of the other two groups (P ＜ 0.05).The integrated optical density (IA) of the experimental group (6 558.2 ± 1 567.1) was significantly higher than that of the other two groups (460.2 ±203.5 and 588.5 ± 241.3,all P ＜0.05).Immunohistochemistry showed that the expression level of ALKBH3 in experimental group (0.26 ± 0.03) were lower than the blank group (0.43 ± 0.07) and control group (0.48 ± 0.06,P ＜ 0.05),the expression level of PCNA protein in the experimental group (0.16 ± 0.02) were lower than the control group (0.28 ± 0.05) and control group (0.31 ± 0.05,all P ＜ 0.05),the expression level of bcl-2 in experimental group (0.31 ±0.05) were lower than the blank group (0.55 ±0.12) and the control group (0.49 ± 0.11,all P ＜ 0.05).Conclusion SiRNA targeting ALKBH3 gene could inhibit obviously the tumor growth by inducing apoptosis in human prostate cancer xenograft in nude mice.Its mechanism of action is possibly related with the downregulation of PCNA and bcl-2 expression.