中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2014年
27期
4-8,20
,共6页
microRNA%视网膜母细胞%肿瘤干细胞%多药耐药%ABCG2(+)/ABCG2(-)
microRNA%視網膜母細胞%腫瘤榦細胞%多藥耐藥%ABCG2(+)/ABCG2(-)
microRNA%시망막모세포%종류간세포%다약내약%ABCG2(+)/ABCG2(-)
microRNA%Retinal mother cells%Cancer stem cells%Multi-drug resistant%ABCG2 (+)/ABCG2 (-)
目的：通过对人视网膜母细胞瘤(RB)肿瘤干细胞(CSCs)多药耐药性的研究，探讨microRNA对肿瘤干细胞多药耐药性的影响。方法通过流式细胞仪分选Y79细胞中的ABCG2(+)与ABCG2(-)细胞，作为ABCG2(+)组与ABCG2(-)组；采用MTT比色实验比较ABCG2(+)组与ABCG2(-)组对抗肿瘤药物长春新碱、依托泊苷和卡铂的敏感性，得到候选microRNA。从标本库中选择RB组织切片，行LNA-原位杂交，并根据耐药性分为高耐药性组与低耐药性组，通过Real time PCR观察候选microRNA在两组中表达差异。结果①分选阳性率：ABCG2(+)分选后(91.7%)显著高于分选前(5.7%)，差异有统计学意义(P<0.05)。②两组耐药性比较：从各个浓度水平上比较，ABCG2(+)细胞的耐药性明显高于ABCG2(-)细胞，差异有统计学意义(P<0.05)。③分选细胞的凋亡率和细胞内药物蓄积比较：ABCG2(+)组的凋亡率明显低于ABCG2(-)组，差异有统计学意义(P<0.05)；ABCG2(+)组的药物蓄积明显低于ABCG2(-)组，差异有统计学意义(P<0.05)。④候选microRNA在不同耐药性RB切片中的表达：高耐药组的microRNA表达率(39.3%)明显高于低耐药组的microRNA表达率低(5.2%)，差异有统计学意义(P<0.05)。结论 ABCG2(+)是Y79细胞系中的耐药细胞群，microRNA与肿瘤干细胞的侵袭性和耐药性密切相关，调控microRNA可以抑制肿瘤干细胞的耐药性，提高其对化疗的敏感性。
目的：通過對人視網膜母細胞瘤(RB)腫瘤榦細胞(CSCs)多藥耐藥性的研究，探討microRNA對腫瘤榦細胞多藥耐藥性的影響。方法通過流式細胞儀分選Y79細胞中的ABCG2(+)與ABCG2(-)細胞，作為ABCG2(+)組與ABCG2(-)組；採用MTT比色實驗比較ABCG2(+)組與ABCG2(-)組對抗腫瘤藥物長春新堿、依託泊苷和卡鉑的敏感性，得到候選microRNA。從標本庫中選擇RB組織切片，行LNA-原位雜交，併根據耐藥性分為高耐藥性組與低耐藥性組，通過Real time PCR觀察候選microRNA在兩組中錶達差異。結果①分選暘性率：ABCG2(+)分選後(91.7%)顯著高于分選前(5.7%)，差異有統計學意義(P<0.05)。②兩組耐藥性比較：從各箇濃度水平上比較，ABCG2(+)細胞的耐藥性明顯高于ABCG2(-)細胞，差異有統計學意義(P<0.05)。③分選細胞的凋亡率和細胞內藥物蓄積比較：ABCG2(+)組的凋亡率明顯低于ABCG2(-)組，差異有統計學意義(P<0.05)；ABCG2(+)組的藥物蓄積明顯低于ABCG2(-)組，差異有統計學意義(P<0.05)。④候選microRNA在不同耐藥性RB切片中的錶達：高耐藥組的microRNA錶達率(39.3%)明顯高于低耐藥組的microRNA錶達率低(5.2%)，差異有統計學意義(P<0.05)。結論 ABCG2(+)是Y79細胞繫中的耐藥細胞群，microRNA與腫瘤榦細胞的侵襲性和耐藥性密切相關，調控microRNA可以抑製腫瘤榦細胞的耐藥性，提高其對化療的敏感性。
목적：통과대인시망막모세포류(RB)종류간세포(CSCs)다약내약성적연구，탐토microRNA대종류간세포다약내약성적영향。방법통과류식세포의분선Y79세포중적ABCG2(+)여ABCG2(-)세포，작위ABCG2(+)조여ABCG2(-)조；채용MTT비색실험비교ABCG2(+)조여ABCG2(-)조대항종류약물장춘신감、의탁박감화잡박적민감성，득도후선microRNA。종표본고중선택RB조직절편，행LNA-원위잡교，병근거내약성분위고내약성조여저내약성조，통과Real time PCR관찰후선microRNA재량조중표체차이。결과①분선양성솔：ABCG2(+)분선후(91.7%)현저고우분선전(5.7%)，차이유통계학의의(P<0.05)。②량조내약성비교：종각개농도수평상비교，ABCG2(+)세포적내약성명현고우ABCG2(-)세포，차이유통계학의의(P<0.05)。③분선세포적조망솔화세포내약물축적비교：ABCG2(+)조적조망솔명현저우ABCG2(-)조，차이유통계학의의(P<0.05)；ABCG2(+)조적약물축적명현저우ABCG2(-)조，차이유통계학의의(P<0.05)。④후선microRNA재불동내약성RB절편중적표체：고내약조적microRNA표체솔(39.3%)명현고우저내약조적microRNA표체솔저(5.2%)，차이유통계학의의(P<0.05)。결론 ABCG2(+)시Y79세포계중적내약세포군，microRNA여종류간세포적침습성화내약성밀절상관，조공microRNA가이억제종류간세포적내약성，제고기대화료적민감성。
Objective To discuss the influence of MicroRNA on multi-drug resistance of cancer stem cell through hu-man retinoblastoma (RB) cancer stem cells (CSCs) multi-drug resistance research. Methods ABCG2 (+) and ABCG2 (-) cells from Y79 cells were sorted by flow cytometry, as the ABCG2 (+)group and the ABCG2 (-) group;the sensitivi-ty of ABCG2 (+) and the ABCG2 (-) cells for vincristine, etoposide and carboplatin were compared by MTT colorimet-ric experiment, and then candidate microRNA was gotten. RB tissue biopsies were chosen from specimens library, then were given LNA-situ hybridization, and divided into high resistance group and low resistance group according to the resistance, microRNA expression differences in the two groups were observed by real-time PCR. Results ①Separating positive rate of ABCG2 (+) after separation (91.7%) was significantly higher than that before the separation (5.7%), the difference was statistically significant (P< 0.05). ②Resistance comparison, ABCG2 resistance of ABCG2 (+) cells was obviously higher than that of ABCG2 (-) cells on various levels, the difference was statistically significant (P<0.05).③The sorting cell apoptosis rate and intracellular drug accumulation comparison, apoptosis rate of ABCG2 (+)group was obviously lower than the ABCG2 (-) group, the difference was statistically significant (P<0.05); the drug accumu-lation of ABCG2 (+) group was significantly lower than the ABCG2 (-) group, the difference was statistically significant (P < 0.05). ④the expression of candidate microRNA in different resistance RB section, microRNA expression rate of high resistance group (39.3%) was significantly higher than low resistance group (5.2%), the difference was statistically significant (P<0.05). Conclusion The ABCG2 (+) is the drug-resistant cell line of Y79 cells, microRNA is closely re-lated to the tumor stem cell invasive and resistance, the microRNA regulating can inhibit the drug resistance of cancer stem cells, improve its sensitivity to chemotherapy.
