中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2015年
1期
47-52
,共6页
石翠娟%王影%于士柱%孙翠云%王虔%徐慧%安同岭%温艳军%徐金玲%刘静%李慧凝
石翠娟%王影%于士柱%孫翠雲%王虔%徐慧%安同嶺%溫豔軍%徐金玲%劉靜%李慧凝
석취연%왕영%우사주%손취운%왕건%서혜%안동령%온염군%서금령%류정%리혜응
胶质瘤%微小RNA-29c%细胞分裂周期蛋白42%细胞增殖
膠質瘤%微小RNA-29c%細胞分裂週期蛋白42%細胞增殖
효질류%미소RNA-29c%세포분렬주기단백42%세포증식
glioma%microRNA-29c%cell division cycle 42%proliferation
目的：探讨microRNA-29c（miR-29c）在胶质瘤中的表达及其对细胞分裂周期蛋白42（CDC42）的调控作用和对细胞增殖的影响。方法：用锁定寡核苷酸原位杂交法和免疫组织化学法检测60例不同级别胶质瘤及10例非肿瘤对照脑组织中miR-29c和CDC42的表达水平，实时荧光定量RT-PCR、Western blot及MTS法分别检测U87MG细胞中miR-29c瞬时表达、CDC42 mRNA和蛋白表达及其对胶质瘤细胞增殖的影响。结果：各级别胶质瘤的miR-29c表达水平均明显低于非肿瘤对照脑组织，且随肿瘤级别升高而显著降低，各组间的差异均有统计学意义（P<0.001）。而CDC42表达水平则呈相反趋势，其表达水平随胶质瘤良恶性级别增加相应升高，除Ⅰ～Ⅱ级组与非肿瘤对照组之间外，其余各组间的差异均有统计学意义（P<0.001）。与对照组相比， miR-29c转染组的CDC42 mRNA（P<0.001）和蛋白表达水平（P<0.01）均明显降低。miR-29c转染组细胞的增殖能力明显低于U87MG空白对照组和Scr转染组（P<0.05）。结论：miR-29c是胶质瘤的抑瘤miRNA，其表达水平可作为评价胶质瘤良恶性级别的重要参考指标。miR-29c在胶质瘤中表达减少解除了其对靶基因CDC42转录后水平的抑制作用，并导致肿瘤细胞无限增殖。表明miR-29c表达异常减少可能是引起胶质瘤发生、发展的关键事件。
目的：探討microRNA-29c（miR-29c）在膠質瘤中的錶達及其對細胞分裂週期蛋白42（CDC42）的調控作用和對細胞增殖的影響。方法：用鎖定寡覈苷痠原位雜交法和免疫組織化學法檢測60例不同級彆膠質瘤及10例非腫瘤對照腦組織中miR-29c和CDC42的錶達水平，實時熒光定量RT-PCR、Western blot及MTS法分彆檢測U87MG細胞中miR-29c瞬時錶達、CDC42 mRNA和蛋白錶達及其對膠質瘤細胞增殖的影響。結果：各級彆膠質瘤的miR-29c錶達水平均明顯低于非腫瘤對照腦組織，且隨腫瘤級彆升高而顯著降低，各組間的差異均有統計學意義（P<0.001）。而CDC42錶達水平則呈相反趨勢，其錶達水平隨膠質瘤良噁性級彆增加相應升高，除Ⅰ～Ⅱ級組與非腫瘤對照組之間外，其餘各組間的差異均有統計學意義（P<0.001）。與對照組相比， miR-29c轉染組的CDC42 mRNA（P<0.001）和蛋白錶達水平（P<0.01）均明顯降低。miR-29c轉染組細胞的增殖能力明顯低于U87MG空白對照組和Scr轉染組（P<0.05）。結論：miR-29c是膠質瘤的抑瘤miRNA，其錶達水平可作為評價膠質瘤良噁性級彆的重要參攷指標。miR-29c在膠質瘤中錶達減少解除瞭其對靶基因CDC42轉錄後水平的抑製作用，併導緻腫瘤細胞無限增殖。錶明miR-29c錶達異常減少可能是引起膠質瘤髮生、髮展的關鍵事件。
목적：탐토microRNA-29c（miR-29c）재효질류중적표체급기대세포분렬주기단백42（CDC42）적조공작용화대세포증식적영향。방법：용쇄정과핵감산원위잡교법화면역조직화학법검측60례불동급별효질류급10례비종류대조뇌조직중miR-29c화CDC42적표체수평，실시형광정량RT-PCR、Western blot급MTS법분별검측U87MG세포중miR-29c순시표체、CDC42 mRNA화단백표체급기대효질류세포증식적영향。결과：각급별효질류적miR-29c표체수평균명현저우비종류대조뇌조직，차수종류급별승고이현저강저，각조간적차이균유통계학의의（P<0.001）。이CDC42표체수평칙정상반추세，기표체수평수효질류량악성급별증가상응승고，제Ⅰ～Ⅱ급조여비종류대조조지간외，기여각조간적차이균유통계학의의（P<0.001）。여대조조상비， miR-29c전염조적CDC42 mRNA（P<0.001）화단백표체수평（P<0.01）균명현강저。miR-29c전염조세포적증식능력명현저우U87MG공백대조조화Scr전염조（P<0.05）。결론：miR-29c시효질류적억류miRNA，기표체수평가작위평개효질류량악성급별적중요삼고지표。miR-29c재효질류중표체감소해제료기대파기인CDC42전록후수평적억제작용，병도치종류세포무한증식。표명miR-29c표체이상감소가능시인기효질류발생、발전적관건사건。
Objective:To investigate microRNA-29c (miR-29c) expression and its relationship with CDC42 in gliomas, as well as to observe its effects on the proliferation of the U87MG glioma cell line. Methods:The expression levels of miR-29c and CDC42 were determined by using locked-oligonucleotide-probe in situ hybridization and immunohistochemistry in 10 cases with nontumor control brain tissues and 60 patients with gliomas of 4 pathological grades. Mature mimics of miR-29c and scrambled sequences were chemically synthesized and then transiently transfected into the U87MG glioma cell line. The miR-29c expression level was quantified by using stem-loop real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The expression levels of CDC42 mRNA and protein, as well as the proliferation capabilities of U87MG, were evaluated by qRT-PCR, Western blot, and MTS assay. Results:Locked-oligonucleotide-probe in situ hybridization showed a downregulation of miR-29c in all glioma samples compared with subjects having nontumor control brain tissues;a continuous decrease was observed as the malignant grade of the tumors increased (P<0.001). CDC42 immunohistochemistry exhibited the opposite pattern. The Labeling index (LI%) value of CDC42 was the highest in the WHO grade IV group. All between-group differences, except for that between the WHO grade I-II and nontumor control groups, were statistically significant (P<0.001). The miR-29c expression levels in miR-29c transcription groups were significantly higher than those in the blank and Scr control groups (P<0.001). Compared with the values for the control groups, the CDC42 mRNA (P<0.001) and protein (P<0.01) levels were significantly decreased in the miR-29c transcription groups. The proliferation capabilities of the U87MG glioma cell line in miR-29c transcription groups were significantly lower than those of the control groups at 48 (P<0.05), 72 (P<0.05), and 96 h (P<0.001) after transient miR-29c transfection. Conclusion:miR-29c is an important tumor-suppressive miRNA that could be used as an important marker to assess the malignant degree of gliomas. The aberrant decrease in miR-29c expression in gliomas resulted in CDC42 upregulation and facilitated glioma cell immortalization. These findings further confirm that miR-29c downregulation may be a key mechanism for glioblastoma tumorigenesis.
