中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2015年
9期
1593-1597
,共5页
刘利平%鲍世韵%张育森%何婉
劉利平%鮑世韻%張育森%何婉
류리평%포세운%장육삼%하완
肝肿瘤%人抗原R%RNA干扰%化疗敏感性%抗药性,多药
肝腫瘤%人抗原R%RNA榦擾%化療敏感性%抗藥性,多藥
간종류%인항원R%RNA간우%화료민감성%항약성,다약
Liver neoplasms%Human antigen R%RNA interference%Chemosensitivity%Drug resistance,multiple
目的：探讨沉默人抗原R（HuR）基因对人肝癌细胞Hep3B多柔比星化疗敏感性的影响及相关机制。方法脂质体包裹靶向HuR基因的短发夹RNA（shRNA）转染Hep3B细胞，real-time PCR和Western blot分别检测沉默HuR基因后HuR、MDR1 mRNA和蛋白的表达，MTT法检测沉默HuR基因后Hep3B细胞对多柔比星化疗的敏感性，流式细胞术（FCM）检测沉默HuR基因后对Hep3B细胞凋亡的影响。结果 HuR shRNA质粒转染Hep3B细胞能有效沉默HuR基因，mRNA和蛋白水平的抑制率分别为82%和75%。与未处理Hep3B细胞（Mock组）比较，沉默HuR基因的Hep3B细胞（siHuR组）对多柔比星的半数抑制浓度（IC50）显著降低[（86.36±6.54）μg/L vs.（318.56±9.89）μg/L， P＜0.05]，细胞凋亡率明显升高[（30.48±5.12）%vs.（8.86±1.35）%，P＜0.05]，MDR1 mRNA和P-gp蛋白表达显著降低，比值分别为0.36±0.08和0.28±0.11，差异均有统计学意义（P＜0.05）。Mock组与转染对照shRNA质粒Hep3B（Control组）的各检测指标均无统计学差异（P＞0.05）。结论沉默HuR基因可能通过抑制耐药蛋白P-gp的表达增强其对多柔比星的化疗敏感性。
目的：探討沉默人抗原R（HuR）基因對人肝癌細胞Hep3B多柔比星化療敏感性的影響及相關機製。方法脂質體包裹靶嚮HuR基因的短髮夾RNA（shRNA）轉染Hep3B細胞，real-time PCR和Western blot分彆檢測沉默HuR基因後HuR、MDR1 mRNA和蛋白的錶達，MTT法檢測沉默HuR基因後Hep3B細胞對多柔比星化療的敏感性，流式細胞術（FCM）檢測沉默HuR基因後對Hep3B細胞凋亡的影響。結果 HuR shRNA質粒轉染Hep3B細胞能有效沉默HuR基因，mRNA和蛋白水平的抑製率分彆為82%和75%。與未處理Hep3B細胞（Mock組）比較，沉默HuR基因的Hep3B細胞（siHuR組）對多柔比星的半數抑製濃度（IC50）顯著降低[（86.36±6.54）μg/L vs.（318.56±9.89）μg/L， P＜0.05]，細胞凋亡率明顯升高[（30.48±5.12）%vs.（8.86±1.35）%，P＜0.05]，MDR1 mRNA和P-gp蛋白錶達顯著降低，比值分彆為0.36±0.08和0.28±0.11，差異均有統計學意義（P＜0.05）。Mock組與轉染對照shRNA質粒Hep3B（Control組）的各檢測指標均無統計學差異（P＞0.05）。結論沉默HuR基因可能通過抑製耐藥蛋白P-gp的錶達增彊其對多柔比星的化療敏感性。
목적：탐토침묵인항원R（HuR）기인대인간암세포Hep3B다유비성화료민감성적영향급상관궤제。방법지질체포과파향HuR기인적단발협RNA（shRNA）전염Hep3B세포，real-time PCR화Western blot분별검측침묵HuR기인후HuR、MDR1 mRNA화단백적표체，MTT법검측침묵HuR기인후Hep3B세포대다유비성화료적민감성，류식세포술（FCM）검측침묵HuR기인후대Hep3B세포조망적영향。결과 HuR shRNA질립전염Hep3B세포능유효침묵HuR기인，mRNA화단백수평적억제솔분별위82%화75%。여미처리Hep3B세포（Mock조）비교，침묵HuR기인적Hep3B세포（siHuR조）대다유비성적반수억제농도（IC50）현저강저[（86.36±6.54）μg/L vs.（318.56±9.89）μg/L， P＜0.05]，세포조망솔명현승고[（30.48±5.12）%vs.（8.86±1.35）%，P＜0.05]，MDR1 mRNA화P-gp단백표체현저강저，비치분별위0.36±0.08화0.28±0.11，차이균유통계학의의（P＜0.05）。Mock조여전염대조shRNA질립Hep3B（Control조）적각검측지표균무통계학차이（P＞0.05）。결론침묵HuR기인가능통과억제내약단백P-gp적표체증강기대다유비성적화료민감성。
Objective To investigate the effect and underlying mechanisms of silencing human antigen R (human antigen R, HuR) gene to the chemotherapy sensitivity of human liver cancer cells Hep3B to doxorubicin. Methods Plasmids of short hairpin RNA (short hairpin RNA, shRNA) targeting HuR gene were transfected into Hep3B cells by Lipofectimine 2000. The mRNA and protein levels of HuR and MDR1 in Hep3B cells after transfection of these plasmids were detected by real-time PCR and Western blot analysis, respectively. The chemosensitivity to doxorubicin and apoptosis rates of Hep3B cells were measured by MTT assay and flow cytometry (FCM). Results Transfection of HuR shRNA plasmids effectively inhibited the mRNA and protein levels of HuR in Hep3B cells by 82%and 75%, respectively. In comparison with untransfected Hep3B cells (Mock group), the half inhibitory concentration (IC50) of Hep3B cells transfected with HuR shRNA plasmids (siHuR group) was significantly lower [(86.36±6.54)μg/L vs. (318.56±9.89)μg/L, P<0.05)]. Meanwhile, the apoptosis rates of siHuR group were increased significantly [(30.48±5.12)%vs. (8.86±1.35)%, P<0.05], and the MDR1 mRNA and P-gp protein expression was significantly lower, with ratio was 0.36±0.08 and 0.28±0.11 (P<0.05). Conclusion Gene silencing of HuR may increase the chemosensitivity of Hep3B to doxorubicin through inhibiting the expression of MDR1 gene and P-gp.
