中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2014年
2期
92-96
,共5页
毛盛勋%何楠%辛林%曾飞%曹家庆
毛盛勛%何楠%辛林%曾飛%曹傢慶
모성훈%하남%신림%증비%조가경
胃肿瘤%miR-224%反义单核苷酸%细胞增殖%细胞凋亡
胃腫瘤%miR-224%反義單覈苷痠%細胞增殖%細胞凋亡
위종류%miR-224%반의단핵감산%세포증식%세포조망
Stomach neoplasms%miR-224%Antisense oligonucleotides%Cell proliferation%Apoptosis
目的 体内外观察miR-224反义核苷酸(ASO)对胃癌细胞增殖和凋亡的影响.方法 采用实时荧光定量PCR法检测120例胃癌组织及其癌旁组织中miR-224 mRNA的表达水平；采用四甲基偶氮唑蓝(MTT)法、克隆形成实验、流式细胞技术及裸鼠体内实验观察miR-224 ASO对SGC7901细胞产生的生物学效应.结果 120例胃癌组织和癌旁组织中miR-224 mRNA的相对表达水平分别为0.28 ±0.07和0.12±0.03,差异有统计学意义(P＜0.05).miR-224 ASO组细胞中miR-224 mRNA 的相对表达水平为0.09 ±0.01,明显低于对照ASO组(0.50 ±0.07,P＜0.05).MTT法检测结果显示,miR-224 ASO组细胞转染后24、48、72 h的细胞生长抑制率分别为53.6％、59.1％和70.1％,对照ASO组分别为12.3％、17.4％和24.7％,差异均有统计学意义(均P＜0.05).克隆形成实验结果显示,miR-224 ASO组的细胞克隆形成率为(5.33±0.74)％,对照ASO组为(33.33±8.38)％,差异有统计学意义(P＜0.05).流式细胞仪检测结果显示,miR-224 ASO组和对照ASO组的细胞凋亡指数分别为(15.68±1.46)％和(3.36±0.88)％,差异有统计学意义(P=0.02).miR-224 ASO组细胞Bcl-2 mRNA和蛋白的相对表达水平分别为1.05 ±0.04和0.21 ±0.03,对照ASO组分别为4.87±0.96和0.88 ±0.09,差异均有统计学意义(均P＜0.01).体内研究显示,接种后30 d,miR-224ASO组裸鼠的肿瘤体积明显低于对照ASO组,差异有统计学意义(P=0.01).结论 miR-224在胃癌组织中表达上调,降低miR-224的表达可有效抑制胃癌细胞的生长,并促进细胞凋亡.miR-224有可能成为胃癌基因表达调控的新靶点.
目的 體內外觀察miR-224反義覈苷痠(ASO)對胃癌細胞增殖和凋亡的影響.方法 採用實時熒光定量PCR法檢測120例胃癌組織及其癌徬組織中miR-224 mRNA的錶達水平；採用四甲基偶氮唑藍(MTT)法、剋隆形成實驗、流式細胞技術及裸鼠體內實驗觀察miR-224 ASO對SGC7901細胞產生的生物學效應.結果 120例胃癌組織和癌徬組織中miR-224 mRNA的相對錶達水平分彆為0.28 ±0.07和0.12±0.03,差異有統計學意義(P＜0.05).miR-224 ASO組細胞中miR-224 mRNA 的相對錶達水平為0.09 ±0.01,明顯低于對照ASO組(0.50 ±0.07,P＜0.05).MTT法檢測結果顯示,miR-224 ASO組細胞轉染後24、48、72 h的細胞生長抑製率分彆為53.6％、59.1％和70.1％,對照ASO組分彆為12.3％、17.4％和24.7％,差異均有統計學意義(均P＜0.05).剋隆形成實驗結果顯示,miR-224 ASO組的細胞剋隆形成率為(5.33±0.74)％,對照ASO組為(33.33±8.38)％,差異有統計學意義(P＜0.05).流式細胞儀檢測結果顯示,miR-224 ASO組和對照ASO組的細胞凋亡指數分彆為(15.68±1.46)％和(3.36±0.88)％,差異有統計學意義(P=0.02).miR-224 ASO組細胞Bcl-2 mRNA和蛋白的相對錶達水平分彆為1.05 ±0.04和0.21 ±0.03,對照ASO組分彆為4.87±0.96和0.88 ±0.09,差異均有統計學意義(均P＜0.01).體內研究顯示,接種後30 d,miR-224ASO組裸鼠的腫瘤體積明顯低于對照ASO組,差異有統計學意義(P=0.01).結論 miR-224在胃癌組織中錶達上調,降低miR-224的錶達可有效抑製胃癌細胞的生長,併促進細胞凋亡.miR-224有可能成為胃癌基因錶達調控的新靶點.
목적 체내외관찰miR-224반의핵감산(ASO)대위암세포증식화조망적영향.방법 채용실시형광정량PCR법검측120례위암조직급기암방조직중miR-224 mRNA적표체수평；채용사갑기우담서람(MTT)법、극륭형성실험、류식세포기술급라서체내실험관찰miR-224 ASO대SGC7901세포산생적생물학효응.결과 120례위암조직화암방조직중miR-224 mRNA적상대표체수평분별위0.28 ±0.07화0.12±0.03,차이유통계학의의(P＜0.05).miR-224 ASO조세포중miR-224 mRNA 적상대표체수평위0.09 ±0.01,명현저우대조ASO조(0.50 ±0.07,P＜0.05).MTT법검측결과현시,miR-224 ASO조세포전염후24、48、72 h적세포생장억제솔분별위53.6％、59.1％화70.1％,대조ASO조분별위12.3％、17.4％화24.7％,차이균유통계학의의(균P＜0.05).극륭형성실험결과현시,miR-224 ASO조적세포극륭형성솔위(5.33±0.74)％,대조ASO조위(33.33±8.38)％,차이유통계학의의(P＜0.05).류식세포의검측결과현시,miR-224 ASO조화대조ASO조적세포조망지수분별위(15.68±1.46)％화(3.36±0.88)％,차이유통계학의의(P=0.02).miR-224 ASO조세포Bcl-2 mRNA화단백적상대표체수평분별위1.05 ±0.04화0.21 ±0.03,대조ASO조분별위4.87±0.96화0.88 ±0.09,차이균유통계학의의(균P＜0.01).체내연구현시,접충후30 d,miR-224ASO조라서적종류체적명현저우대조ASO조,차이유통계학의의(P=0.01).결론 miR-224재위암조직중표체상조,강저miR-224적표체가유효억제위암세포적생장,병촉진세포조망.miR-224유가능성위위암기인표체조공적신파점.
Objective To observe the effects of miR-224 antisense oligonucleotide (ASO) on the proliferation and apoptosis of gastric cancer cells in vitro and vivo.Methods The expression of miR-224 inthe cancer tissues and their adjacent tissues in 120 gastric cancer patients were detected by real-time quantitative PCR.The biological effects of miR-224 ASO on human gastric cancer SGC7901 cells was assessed by MTT assay,clone formation assay,flow cytometry and in vivo experiment in nude mice.Results Compared with the control group (0.50 ± 0.07),miR-224 ASO significantly reduced the miR-224 mRNA expression in the cancer patients (0.09 ± 0.01,P ＜ 0.05).MTT assay results showed that the survival rate of gastric cells at 24 h,48 h and 72 h was 53.6％,59.1％ and 70.1％ in the miR-224 ASO group,and 12.3％,17.4％ and 24.7％,respectively,in the control group (P ＜ 0.05 for all).Clone formation assay revealed that clone formation rate in the miR-224 ASO group was (5.33 ± 0.74) ％,significantly lower than the (33.33 ± 8.38) ％ in the control group (P ＜ 0.05).Flow cytometry indicated that the apoptotic index was (15.68 ±1.46)％ in the miR-224 ASO group and (3.36 ±0.88)％ in the control group (P＜0.01).In addition,the expressions of Bcl2 mRNA and protein were 1.05-±0.04 and 0.21 ±0.03 in the miR-224 ASO group,significantly lower than that in the control group (4.87 ±0.96 and 0.88 ±0.09,P ＜0.01).The in vivo study further showed that the tumor volume in the experimental group is significantly smaller than that in the control group (P =0.01).Conclusions MiR-224 is overexpressed in human gastric cancer.Reducing the expression of miR-224 can effectively inhibit the growth and promote apoptosis of gastric cancer cells,miR-224 may become a new target for the regulation of gene expression in gastric cancer.
