中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2012年
9期
664-668
,共5页
陈佳萍%刘瑶%胡志斌%沈洪兵
陳佳萍%劉瑤%鬍誌斌%瀋洪兵
진가평%류요%호지빈%침홍병
微小RNA-30c%单核苷酸多态性%转录,遗传%肺肿瘤
微小RNA-30c%單覈苷痠多態性%轉錄,遺傳%肺腫瘤
미소RNA-30c%단핵감산다태성%전록,유전%폐종류
MiR-3Oc%Single nucleotide polymorphism%Transcrition,genetic%Lung neoplasms
目的 研究不同细胞和肺癌组织中miR-30c侧翼区单核苷酸多态性rs928508(A/G)对初级miR-30c(pri-miR-30c)、前体miR-30c(pre-miR-30c)、成熟miR-30c的表达水平以及miR-30c成熟进程的影响.方法 构建含有不同基因型miR-30c侧翼区的报告基因表达载体pGL3-promoter-miR-30c-A和pGL3-promoter-miR-30c-G,分别与内参质粒pRL-SV40共转染于人肺癌A549细胞、人胚肾293A细胞和中国仓鼠卵巢细胞(CHO细胞)后,进行荧光素酶活性分析.采用Taqman基因分型技术对50例肺癌组织进行基因分型,以实时定量PCR法检测不同基因型组织标本中pri-miR-30c、pre-miR-30c、成熟miR-30c以及miR-30c宿主基因核转录因子Y(NFYC)的转录水平.结果 在A549、293A和CHO细胞中,pGL3-promoter-miR-30c-A组荧光素酶的活性与pGL3-promoter-miR-30c-G组比较,差异均无统计学意义(均P＞0.05).基因分型的结果显示,50例肺癌组织中,AA基因型17例,AG基因型21例,GG基因型12例.在rs928508 AG/GG基因型肺癌组织中,pre-miR-30c和成熟miR-30c的相对表达水平分别为1.306±0.878和0.384±0.227,均显著低于rs928508 AA基因型组织的表达水平(分别为2.103±1.173和0.588±0.307,P=0.009和P=0.011).而pri-miR-30c和NFYC基因在各基因型肺癌组织中的相对表达量差异均无统计学意义(P =0.335和P=0.393).结论 在肺癌组织中,miR-30c侧翼区的单核苷酸多态性rs928508(A/G)可以影响p6-miR40c→pre-miR-30c→成熟miR-30c的转化进程,但不能影响pri-miR-30c合成的转录进程.
目的 研究不同細胞和肺癌組織中miR-30c側翼區單覈苷痠多態性rs928508(A/G)對初級miR-30c(pri-miR-30c)、前體miR-30c(pre-miR-30c)、成熟miR-30c的錶達水平以及miR-30c成熟進程的影響.方法 構建含有不同基因型miR-30c側翼區的報告基因錶達載體pGL3-promoter-miR-30c-A和pGL3-promoter-miR-30c-G,分彆與內參質粒pRL-SV40共轉染于人肺癌A549細胞、人胚腎293A細胞和中國倉鼠卵巢細胞(CHO細胞)後,進行熒光素酶活性分析.採用Taqman基因分型技術對50例肺癌組織進行基因分型,以實時定量PCR法檢測不同基因型組織標本中pri-miR-30c、pre-miR-30c、成熟miR-30c以及miR-30c宿主基因覈轉錄因子Y(NFYC)的轉錄水平.結果 在A549、293A和CHO細胞中,pGL3-promoter-miR-30c-A組熒光素酶的活性與pGL3-promoter-miR-30c-G組比較,差異均無統計學意義(均P＞0.05).基因分型的結果顯示,50例肺癌組織中,AA基因型17例,AG基因型21例,GG基因型12例.在rs928508 AG/GG基因型肺癌組織中,pre-miR-30c和成熟miR-30c的相對錶達水平分彆為1.306±0.878和0.384±0.227,均顯著低于rs928508 AA基因型組織的錶達水平(分彆為2.103±1.173和0.588±0.307,P=0.009和P=0.011).而pri-miR-30c和NFYC基因在各基因型肺癌組織中的相對錶達量差異均無統計學意義(P =0.335和P=0.393).結論 在肺癌組織中,miR-30c側翼區的單覈苷痠多態性rs928508(A/G)可以影響p6-miR40c→pre-miR-30c→成熟miR-30c的轉化進程,但不能影響pri-miR-30c閤成的轉錄進程.
목적 연구불동세포화폐암조직중miR-30c측익구단핵감산다태성rs928508(A/G)대초급miR-30c(pri-miR-30c)、전체miR-30c(pre-miR-30c)、성숙miR-30c적표체수평이급miR-30c성숙진정적영향.방법 구건함유불동기인형miR-30c측익구적보고기인표체재체pGL3-promoter-miR-30c-A화pGL3-promoter-miR-30c-G,분별여내삼질립pRL-SV40공전염우인폐암A549세포、인배신293A세포화중국창서란소세포(CHO세포)후,진행형광소매활성분석.채용Taqman기인분형기술대50례폐암조직진행기인분형,이실시정량PCR법검측불동기인형조직표본중pri-miR-30c、pre-miR-30c、성숙miR-30c이급miR-30c숙주기인핵전록인자Y(NFYC)적전록수평.결과 재A549、293A화CHO세포중,pGL3-promoter-miR-30c-A조형광소매적활성여pGL3-promoter-miR-30c-G조비교,차이균무통계학의의(균P＞0.05).기인분형적결과현시,50례폐암조직중,AA기인형17례,AG기인형21례,GG기인형12례.재rs928508 AG/GG기인형폐암조직중,pre-miR-30c화성숙miR-30c적상대표체수평분별위1.306±0.878화0.384±0.227,균현저저우rs928508 AA기인형조직적표체수평(분별위2.103±1.173화0.588±0.307,P=0.009화P=0.011).이pri-miR-30c화NFYC기인재각기인형폐암조직중적상대표체량차이균무통계학의의(P =0.335화P=0.393).결론 재폐암조직중,miR-30c측익구적단핵감산다태성rs928508(A/G)가이영향p6-miR40c→pre-miR-30c→성숙miR-30c적전화진정,단불능영향pri-miR-30c합성적전록진정.
Objective To investigate the effect of a common polymorphism rs928508 (A/G) in flanking region of miR-30c on the expression of pri,pre and mature miR-30c,and discuss the effect of this polymorphism on the maturing process of miR-30c in lung carcinoma.Methods The pGL3-promoter-miR-30c-A and pGL3-promoter-miR-30c-G lueiferase plasmids were created containing A or G allele of miR-30c flanking region.Taqman assay was used to genotype rs928508 polymorphism in 50 lung cancer tissues.RTPCR was performed to determine the expression of pri-miR-30c,pre-miR-30c,mature miR-30c and miR-30c host gene NFYC in the 50 lung cancer tissues.Results The luciferase expression level of the pGL3-promoter-miR-30c-A construct group was not significantly different compared with that in the the pGL3-promoter-miR-30c-G construct group (A549 cells,P =0.758 ; 293A cells,P =0.554 ; CHO cells,P =0.175).The results demonstrated that rs928508 (A/G) variant had no effect on the transcriptional regulation of pri-miR-30c.In the genotype-phenotype collection analysis of the 50 lung cancer tissues,the expression of pre-miR-30c and mature miR-30c for rs928508 AG/GG genotypes showed significantly lower levels compared with those in the AA genotype (P =0.009,P =0.011).However,the expression of primiR-30c showed no significant difference between AG/GG genotypes and AA genotype.Similarly,the expression of host NFYC gene was correlated with pri-miR-30c,showed no significant difference between AG/GG genotypes and AA genotype.Conclusion The rs928508(A/G) polymorphism in flanking region of miR-30c could influence the processing from pri-miR-30c to mature miR-30c,but does not influence the transcription of pri-miR-30c.
