浙江中西医结合杂志
浙江中西醫結閤雜誌
절강중서의결합잡지
ZHEJIANG JOURNAL OF INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE
2015年
8期
720-722,726
,共4页
宫颈癌%miR-26b%Mcl-1%Hela%顺铂
宮頸癌%miR-26b%Mcl-1%Hela%順鉑
궁경암%miR-26b%Mcl-1%Hela%순박
cervical cancer%miR-26b%Mcl-1%Hela%cisplatin
目的：观察microRNA-26b（miR-26b）联合顺铂对宫颈癌细胞的杀伤效果，并探讨其作用机制。方法荧光定量PCR方法检测人正常宫颈上皮细胞系CRL-2614及宫颈癌细胞系Hela、Si-Ha和C-33a的miR-26b表达水平。MTT法检测顺铂单独治疗及联合miR-26b治疗对宫颈癌细胞系Hela的杀伤活性。利用生物信息学及Western blot方法验证miR-26b是否调节Hela细胞Mcl-1表达。构建Mcl-1真核表达载体，MTT法检测Mcl-1表达载体转染对miR-26b联合顺铂杀伤Hela细胞疗效的影响。结果宫颈癌细胞系miR-26b表达水平：Hela细胞为（0.21±0.04），SiHa细胞为（0.42±0.03），C-33a细胞为（0.33±0.03），显著低于正常宫颈上皮细胞系CRL-2614的（1.00±0.05）（P﹤0.05）。miR-26b联合1μmol/L顺铂治疗组对Hela细胞的杀伤活性[细胞活力抑制率为（52.6±6.9）％]显著高于1μmol/L顺铂单治疗组[细胞活力抑制率为（6.7±3.5）％]。miR-26b转染后，Hela细胞Mcl-1蛋白表达水平下降。miR-26b联合1μmol/L顺铂在Mcl-1表达载体转染后对Hela细胞的杀伤活性[细胞活力抑制率为（19.6±6.7）％]显著低于未转染Mcl-1表达载体的miR-26b联合1μmol/L顺铂组[细胞活力抑制率为（55.7±7.6）％]。结论 MiR-26b通过靶向于Mcl-1增强顺铂对宫颈癌细胞系Hela的杀伤活性。
目的：觀察microRNA-26b（miR-26b）聯閤順鉑對宮頸癌細胞的殺傷效果，併探討其作用機製。方法熒光定量PCR方法檢測人正常宮頸上皮細胞繫CRL-2614及宮頸癌細胞繫Hela、Si-Ha和C-33a的miR-26b錶達水平。MTT法檢測順鉑單獨治療及聯閤miR-26b治療對宮頸癌細胞繫Hela的殺傷活性。利用生物信息學及Western blot方法驗證miR-26b是否調節Hela細胞Mcl-1錶達。構建Mcl-1真覈錶達載體，MTT法檢測Mcl-1錶達載體轉染對miR-26b聯閤順鉑殺傷Hela細胞療效的影響。結果宮頸癌細胞繫miR-26b錶達水平：Hela細胞為（0.21±0.04），SiHa細胞為（0.42±0.03），C-33a細胞為（0.33±0.03），顯著低于正常宮頸上皮細胞繫CRL-2614的（1.00±0.05）（P﹤0.05）。miR-26b聯閤1μmol/L順鉑治療組對Hela細胞的殺傷活性[細胞活力抑製率為（52.6±6.9）％]顯著高于1μmol/L順鉑單治療組[細胞活力抑製率為（6.7±3.5）％]。miR-26b轉染後，Hela細胞Mcl-1蛋白錶達水平下降。miR-26b聯閤1μmol/L順鉑在Mcl-1錶達載體轉染後對Hela細胞的殺傷活性[細胞活力抑製率為（19.6±6.7）％]顯著低于未轉染Mcl-1錶達載體的miR-26b聯閤1μmol/L順鉑組[細胞活力抑製率為（55.7±7.6）％]。結論 MiR-26b通過靶嚮于Mcl-1增彊順鉑對宮頸癌細胞繫Hela的殺傷活性。
목적：관찰microRNA-26b（miR-26b）연합순박대궁경암세포적살상효과，병탐토기작용궤제。방법형광정량PCR방법검측인정상궁경상피세포계CRL-2614급궁경암세포계Hela、Si-Ha화C-33a적miR-26b표체수평。MTT법검측순박단독치료급연합miR-26b치료대궁경암세포계Hela적살상활성。이용생물신식학급Western blot방법험증miR-26b시부조절Hela세포Mcl-1표체。구건Mcl-1진핵표체재체，MTT법검측Mcl-1표체재체전염대miR-26b연합순박살상Hela세포료효적영향。결과궁경암세포계miR-26b표체수평：Hela세포위（0.21±0.04），SiHa세포위（0.42±0.03），C-33a세포위（0.33±0.03），현저저우정상궁경상피세포계CRL-2614적（1.00±0.05）（P﹤0.05）。miR-26b연합1μmol/L순박치료조대Hela세포적살상활성[세포활력억제솔위（52.6±6.9）％]현저고우1μmol/L순박단치료조[세포활력억제솔위（6.7±3.5）％]。miR-26b전염후，Hela세포Mcl-1단백표체수평하강。miR-26b연합1μmol/L순박재Mcl-1표체재체전염후대Hela세포적살상활성[세포활력억제솔위（19.6±6.7）％]현저저우미전염Mcl-1표체재체적miR-26b연합1μmol/L순박조[세포활력억제솔위（55.7±7.6）％]。결론 MiR-26b통과파향우Mcl-1증강순박대궁경암세포계Hela적살상활성。
Objective To investigate the effect of microRNA-26b(miR-26b) on cisplatin therapy in treatment of cervical cancer in vitro, and to explore the underlying mechanism. Methods The miR-26b level in normal cervi-cal cell line CRL-2614 and cervical cancer cell lines Hela, SiHa and C-33a were detected by using RT-qPCR. MTT assay was performed to measure the growth inhibition capacity of miR-26b plus cisplatin in Hela cells. Bioinformatics and western blot were performed to determine whether the expression of Mcl-1 in Hela cells was regulated by miR-26b. A Mcl-1 expression vector was constructed to detect the role of Mcl-1 vector toward miR-26b plus cisplatin-inducing cytotoxicity in Hela cells by MTT assay. Results A down-regulation of miR-26b was found in cervical cancer cell lines Hela(0.21±0.04), SiHa(0.42±0.03), and C-33a(0.33±0.03) compared with that in normal cervical cell line CRL-2614 (1.00 ±0.05). The miR-26b plus 1mol/L cisplatin group showed a higher growth inhibition(52.6%±6.9%) than 1μmol/L cisplation group (6.7%±3.5%) in Hela cells. The expression of Mcl-1 at protein level was down-regulated after miR-26b transfection. The growth inhibition of Hela cells treated with miR-26b plus cisplatin was significantly decreased after transfection of Mcl-1 expression vector. Conclusion miR-26b sensitizes cisplatin-induced cytotoxicity by targeting Mcl-1 in cervical cancer.
