中华泌尿外科杂志
中華泌尿外科雜誌
중화비뇨외과잡지
CHINESE JOURNAL OF UROLOGY
2014年
5期
373-377
,共5页
李瑜%李永强%梁卫民%万彦青%谢松强
李瑜%李永彊%樑衛民%萬彥青%謝鬆彊
리유%리영강%량위민%만언청%사송강
槲皮素%顺铂%细胞周期%凋亡%增效
槲皮素%順鉑%細胞週期%凋亡%增效
곡피소%순박%세포주기%조망%증효
Quercetin%Cisplatin%Cell cycle%Apoptosis%Synergism
目的 探讨槲皮素对顺铂抑制前列腺癌细胞的增强作用. 方法 2013年3月选取前列腺癌LNCaP细胞和PC3细胞接种于96孔板12 h.将两种细胞各分为4组:槲皮素组,分别加入10、20、40、80、160 μmol/L槲皮素;顺铂组,分别加入0.01、0.05、0.10、1.00、10.00 μmol/L顺铂;联用组,分别加入20μmol/L槲皮素和0.01、0.05、0.10、1.00、10.00μmol/L顺铂;阴性对照组.培养48 h后,采用噻唑蓝法检测LNCaP细胞和PC3细胞增殖情况.LNCaP细胞和PC3细胞分别加入槲皮素(20 μmol/L)、顺铂(0.05 μmol/L)、槲皮素(20 μmol/L)+顺铂(0.05 μmol/L)培养48 h后,经碘化丙啶及Annexin V-FITC染色后应用流式细胞仪分别检测细胞周期变化情况及凋亡率;蛋白质印迹法检测两种细胞中caspase 8、caspase 9、caspase 3、Bcl-2、Bax等蛋白的表达水平. 结果 顺铂组、槲皮素组的LNCaP细胞及PC3细胞半数抑制浓度(50％ inhibited concentration,ICS0)分别为(0.99±0.13) μmol/L及(0.75±0.09)μmol/L、(91.60±6.10) μmol/L及(72.90±4.70) μmol/L,联用组IC50分别为(0.11±0.06) μmol/L及(0.07±0.02) μmol/L,与槲皮素组和顺铂组比较差异均有统计学意义(P＜0.05).顺铂组、联用组的LNCaP细胞及PC3细胞中S期细胞的比例分别为(22.4±2.7)％及(31.2±2.4)％、(20.1±1.6)％及(31.0±2.5)％,与阴性对照组比较差异均有统计学意义(P＜0.05);槲皮素组与阴性对照组相比较差异无统计学意义(P＞0.05).顺铂组、联用组的LNCaP细胞及PC3细胞的凋亡率分别为(14.8±1.9)％及(39.6±3.1)％、(11.5±1.2)％及(34.1±3.3)％,与阴性对照组比较差异均有统计学意义(P＜0.05);槲皮素组与阴性对照组比较差异无统计学意义(P＞0.05).蛋白质印迹法检测发现槲皮素组、顺铂组、联用组中caspase-3、caspase-8、caspase-9均轻度活化、Bax和p21表达均上调、Bcl-2和CDK2表达均下调、Cytochrome c被释放到胞质中,且顺铂组、联用组的作用均强于槲皮素组,差异均有统计学意义(P＜0.05). 结论 槲皮素能明显增强顺铂对前列腺癌LNCaP细胞和PC3细胞的抑制作用.
目的 探討槲皮素對順鉑抑製前列腺癌細胞的增彊作用. 方法 2013年3月選取前列腺癌LNCaP細胞和PC3細胞接種于96孔闆12 h.將兩種細胞各分為4組:槲皮素組,分彆加入10、20、40、80、160 μmol/L槲皮素;順鉑組,分彆加入0.01、0.05、0.10、1.00、10.00 μmol/L順鉑;聯用組,分彆加入20μmol/L槲皮素和0.01、0.05、0.10、1.00、10.00μmol/L順鉑;陰性對照組.培養48 h後,採用噻唑藍法檢測LNCaP細胞和PC3細胞增殖情況.LNCaP細胞和PC3細胞分彆加入槲皮素(20 μmol/L)、順鉑(0.05 μmol/L)、槲皮素(20 μmol/L)+順鉑(0.05 μmol/L)培養48 h後,經碘化丙啶及Annexin V-FITC染色後應用流式細胞儀分彆檢測細胞週期變化情況及凋亡率;蛋白質印跡法檢測兩種細胞中caspase 8、caspase 9、caspase 3、Bcl-2、Bax等蛋白的錶達水平. 結果 順鉑組、槲皮素組的LNCaP細胞及PC3細胞半數抑製濃度(50％ inhibited concentration,ICS0)分彆為(0.99±0.13) μmol/L及(0.75±0.09)μmol/L、(91.60±6.10) μmol/L及(72.90±4.70) μmol/L,聯用組IC50分彆為(0.11±0.06) μmol/L及(0.07±0.02) μmol/L,與槲皮素組和順鉑組比較差異均有統計學意義(P＜0.05).順鉑組、聯用組的LNCaP細胞及PC3細胞中S期細胞的比例分彆為(22.4±2.7)％及(31.2±2.4)％、(20.1±1.6)％及(31.0±2.5)％,與陰性對照組比較差異均有統計學意義(P＜0.05);槲皮素組與陰性對照組相比較差異無統計學意義(P＞0.05).順鉑組、聯用組的LNCaP細胞及PC3細胞的凋亡率分彆為(14.8±1.9)％及(39.6±3.1)％、(11.5±1.2)％及(34.1±3.3)％,與陰性對照組比較差異均有統計學意義(P＜0.05);槲皮素組與陰性對照組比較差異無統計學意義(P＞0.05).蛋白質印跡法檢測髮現槲皮素組、順鉑組、聯用組中caspase-3、caspase-8、caspase-9均輕度活化、Bax和p21錶達均上調、Bcl-2和CDK2錶達均下調、Cytochrome c被釋放到胞質中,且順鉑組、聯用組的作用均彊于槲皮素組,差異均有統計學意義(P＜0.05). 結論 槲皮素能明顯增彊順鉑對前列腺癌LNCaP細胞和PC3細胞的抑製作用.
목적 탐토곡피소대순박억제전렬선암세포적증강작용. 방법 2013년3월선취전렬선암LNCaP세포화PC3세포접충우96공판12 h.장량충세포각분위4조:곡피소조,분별가입10、20、40、80、160 μmol/L곡피소;순박조,분별가입0.01、0.05、0.10、1.00、10.00 μmol/L순박;련용조,분별가입20μmol/L곡피소화0.01、0.05、0.10、1.00、10.00μmol/L순박;음성대조조.배양48 h후,채용새서람법검측LNCaP세포화PC3세포증식정황.LNCaP세포화PC3세포분별가입곡피소(20 μmol/L)、순박(0.05 μmol/L)、곡피소(20 μmol/L)+순박(0.05 μmol/L)배양48 h후,경전화병정급Annexin V-FITC염색후응용류식세포의분별검측세포주기변화정황급조망솔;단백질인적법검측량충세포중caspase 8、caspase 9、caspase 3、Bcl-2、Bax등단백적표체수평. 결과 순박조、곡피소조적LNCaP세포급PC3세포반수억제농도(50％ inhibited concentration,ICS0)분별위(0.99±0.13) μmol/L급(0.75±0.09)μmol/L、(91.60±6.10) μmol/L급(72.90±4.70) μmol/L,련용조IC50분별위(0.11±0.06) μmol/L급(0.07±0.02) μmol/L,여곡피소조화순박조비교차이균유통계학의의(P＜0.05).순박조、련용조적LNCaP세포급PC3세포중S기세포적비례분별위(22.4±2.7)％급(31.2±2.4)％、(20.1±1.6)％급(31.0±2.5)％,여음성대조조비교차이균유통계학의의(P＜0.05);곡피소조여음성대조조상비교차이무통계학의의(P＞0.05).순박조、련용조적LNCaP세포급PC3세포적조망솔분별위(14.8±1.9)％급(39.6±3.1)％、(11.5±1.2)％급(34.1±3.3)％,여음성대조조비교차이균유통계학의의(P＜0.05);곡피소조여음성대조조비교차이무통계학의의(P＞0.05).단백질인적법검측발현곡피소조、순박조、련용조중caspase-3、caspase-8、caspase-9균경도활화、Bax화p21표체균상조、Bcl-2화CDK2표체균하조、Cytochrome c피석방도포질중,차순박조、련용조적작용균강우곡피소조,차이균유통계학의의(P＜0.05). 결론 곡피소능명현증강순박대전렬선암LNCaP세포화PC3세포적억제작용.
Objective To study the synergistic antitumor effects of quercetin and cisplatin in human prostate cancer PC3 cells and LNCaP cells.Methods Twelve h after PC3 cells or LNCaP cells were seeded,different dose of quercetin (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L,160 μmol/L) or cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L),or quercetin (20 μmol/L) + cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L) were added for48 h and then the antiproliferative effects were detected with MTT assay.After incubated with quercetin (20 μmol/L) or cisplatin (0.05 μ mol/L),or quercetin (20 μ mol/L) + cisplatin (0.05 μmol/L) for48 h,cell cycle distribution and apoptosis of PC3 cells or LNCaP cells were detected by flow cytometer,PI and Annexin V staining.Protein expression was detected by Western blotting.Results After treatment with quercetin or cisplatin alone,the IC50 were (0.99 ± 0.13) μmol/L,(0.75 ± 0.09) μmol/L and (91.60 ± 6.10) μ mol/L,(72.90±4.70) μ mol/L for LNCaP cells or PC3 cells,respectively;The IC50 were (0.11±0.06)μ mol/L,(0.07±0.02) μmol/L for quercetin + cisplatin treatment (Compared with quercetin,P＜0.01 ;Compared with cisplatin,P＜0.05.After treatment with cisplatin or quercetin + cisplatin for 48 h,the S phase percent of LNCaP cells or PC3 cells were (22.4±2.7)％,(31.2±2.4)％ and (20.1±1.6)％,(31.0±2.5)％,respectively,(Compared with control,P＜0.05,however,treatment with quercetin alone has no significant difference (Compared with control,P＞0.05).After treatment with cisplatin or quercetin + cisplatin for 48 h,the apoptotic percent of LNCaP cells or PC3 cells were (14.8 ± 1.9) ％,(39.6 ± 3.1) ％ and (11.5± 1.2) ％,(34.1 ±3.3) ％,respectively,(compared with control,P ＜ 0.05,however,treatment with quercetin alone had no significant difference (compared with control,P＞0.05).After treatment with quercetin alone for 48 h,the activation of caspase-3,caspase-8 and caspase-9 were slightly increased,the expressions of Bax and p21 were up-regulated,the expressions of Bcl-2 and CDK2 were down-regulated.Furthermore,these effect of cisplatin and quercetin + cisplatin were significantly enhanced (compared with quercetin,P＜0.05;compared with quercetin,P＜0.01,respectively.Conclusions The combination modality with quercetin and cisplatin has a better treatment effect in vitro not only in androgen-dependent LNCaP cells but also in androgen-independent PC3 cells.
