目的 观察亚砷酸和顺铂对骨肉瘤MG-63细胞生长的抑制作用.方法 采用细胞培养的方法,体外培养人骨肉瘤株MG-63细胞.应用倒置相差显微镜、透射电镜、四氮唑盐(MTT)比色、流式细胞仪检测等方法,观察亚砷酸和顺铂[各加入100μl(1 g/L)]对骨肉瘤MG-63细胞生长的抑制作用和对细胞凋亡、细胞周期的影响.结果 相差显微镜下,对照组MG-63细胞贴壁生长良好,细胞核呈圆形,细胞呈上皮样细胞排列;亚砷酸组MG-63细胞排列无规则,有细胞死亡;顺铂组MG-63细胞多变小,变圆,胞质的空泡化明显.电镜下,对照组MG-63细胞表面可见球形质膜突起,线粒体嵴粗大,双层核膜结构清晰;亚砷酸组MG-63细胞表面的球形突起消失,线粒体嵴变细,细胞核缩小,凋亡细胞核膜清晰;顺铂组MG-63细胞表面球形突起脱离,细胞核膜增厚,核内染色质形成细纱状.亚砷酸和顺铂均能抑制MG-63细胞生长,抑制率(%)组间比较差异有统计学意义(F=78.859,P<0.05);在2、4、8、16、32、64、128 h,亚砷酸(56.31±0.03、70.00±0.06、79.84±0.03、87.31±0.13、84.70±0.09、90.68±0.06、91.18±0.05)和顺铂(7.55±0.15、15.70±0.17、30.72±0.07、49.80±0.05、45.11±0.13、61.62±0.08、93.80±0.12)对细胞生长的抑制率与对照组(2.03±0.07、2.78±0.08、3.11±0.01、5.67±0.04、12.23±0.04、18.65±0.04、24.45±0.04)比较明显升高(P均<0.05);但与顺铂比较,亚砷酸起效快,在2～32 h抑制率明显高于同时间顺铂(P均<0.05).亚砷酸和顺铂均可导致MG-63细胞凋亡,凋亡率组间比较差异有统计学意义(F=13.317,P<0.05);亚砷酸在24、36、48 h(20.5±3.78、45.76±9.90、25.16±15.41),顺铂在24、36、48 h(12.55±1.51、18.85±3.40、12.37±5.43),MG-63细胞凋亡率(%)明显高于同时间对照组(6.57±1.48、8.03±2.08、6.54±1.30,P<0.05或<0.01).亚砷酸和顺铂均对MG-63细胞周期G1、S、G2/M期有抑制作用,抑制率组间比较差异有统计学意义(F值分别为54.579、43.429、21.795,P<0.05或<0.01);对G1期抑制率(%)亚砷酸(78.26±5.24)和顺铂(80.48±2.81)与对照组(57.49±6.65)比较明显升高(P<0.05或<0.01).结论 亚砷酸和顺铂都可抑制骨肉瘤MG-63细胞生长,诱导细胞凋亡;亚砷酸作用快于顺铂,亚砷酸和顺铂对MG-63细胞的抑制作用发生在细胞周期中的G1期.
目的 觀察亞砷痠和順鉑對骨肉瘤MG-63細胞生長的抑製作用.方法 採用細胞培養的方法,體外培養人骨肉瘤株MG-63細胞.應用倒置相差顯微鏡、透射電鏡、四氮唑鹽(MTT)比色、流式細胞儀檢測等方法,觀察亞砷痠和順鉑[各加入100μl(1 g/L)]對骨肉瘤MG-63細胞生長的抑製作用和對細胞凋亡、細胞週期的影響.結果 相差顯微鏡下,對照組MG-63細胞貼壁生長良好,細胞覈呈圓形,細胞呈上皮樣細胞排列;亞砷痠組MG-63細胞排列無規則,有細胞死亡;順鉑組MG-63細胞多變小,變圓,胞質的空泡化明顯.電鏡下,對照組MG-63細胞錶麵可見毬形質膜突起,線粒體嵴粗大,雙層覈膜結構清晰;亞砷痠組MG-63細胞錶麵的毬形突起消失,線粒體嵴變細,細胞覈縮小,凋亡細胞覈膜清晰;順鉑組MG-63細胞錶麵毬形突起脫離,細胞覈膜增厚,覈內染色質形成細紗狀.亞砷痠和順鉑均能抑製MG-63細胞生長,抑製率(%)組間比較差異有統計學意義(F=78.859,P<0.05);在2、4、8、16、32、64、128 h,亞砷痠(56.31±0.03、70.00±0.06、79.84±0.03、87.31±0.13、84.70±0.09、90.68±0.06、91.18±0.05)和順鉑(7.55±0.15、15.70±0.17、30.72±0.07、49.80±0.05、45.11±0.13、61.62±0.08、93.80±0.12)對細胞生長的抑製率與對照組(2.03±0.07、2.78±0.08、3.11±0.01、5.67±0.04、12.23±0.04、18.65±0.04、24.45±0.04)比較明顯升高(P均<0.05);但與順鉑比較,亞砷痠起效快,在2～32 h抑製率明顯高于同時間順鉑(P均<0.05).亞砷痠和順鉑均可導緻MG-63細胞凋亡,凋亡率組間比較差異有統計學意義(F=13.317,P<0.05);亞砷痠在24、36、48 h(20.5±3.78、45.76±9.90、25.16±15.41),順鉑在24、36、48 h(12.55±1.51、18.85±3.40、12.37±5.43),MG-63細胞凋亡率(%)明顯高于同時間對照組(6.57±1.48、8.03±2.08、6.54±1.30,P<0.05或<0.01).亞砷痠和順鉑均對MG-63細胞週期G1、S、G2/M期有抑製作用,抑製率組間比較差異有統計學意義(F值分彆為54.579、43.429、21.795,P<0.05或<0.01);對G1期抑製率(%)亞砷痠(78.26±5.24)和順鉑(80.48±2.81)與對照組(57.49±6.65)比較明顯升高(P<0.05或<0.01).結論 亞砷痠和順鉑都可抑製骨肉瘤MG-63細胞生長,誘導細胞凋亡;亞砷痠作用快于順鉑,亞砷痠和順鉑對MG-63細胞的抑製作用髮生在細胞週期中的G1期.
목적 관찰아신산화순박대골육류MG-63세포생장적억제작용.방법 채용세포배양적방법,체외배양인골육류주MG-63세포.응용도치상차현미경、투사전경、사담서염(MTT)비색、류식세포의검측등방법,관찰아신산화순박[각가입100μl(1 g/L)]대골육류MG-63세포생장적억제작용화대세포조망、세포주기적영향.결과 상차현미경하,대조조MG-63세포첩벽생장량호,세포핵정원형,세포정상피양세포배렬;아신산조MG-63세포배렬무규칙,유세포사망;순박조MG-63세포다변소,변원,포질적공포화명현.전경하,대조조MG-63세포표면가견구형질막돌기,선립체척조대,쌍층핵막결구청석;아신산조MG-63세포표면적구형돌기소실,선립체척변세,세포핵축소,조망세포핵막청석;순박조MG-63세포표면구형돌기탈리,세포핵막증후,핵내염색질형성세사상.아신산화순박균능억제MG-63세포생장,억제솔(%)조간비교차이유통계학의의(F=78.859,P<0.05);재2、4、8、16、32、64、128 h,아신산(56.31±0.03、70.00±0.06、79.84±0.03、87.31±0.13、84.70±0.09、90.68±0.06、91.18±0.05)화순박(7.55±0.15、15.70±0.17、30.72±0.07、49.80±0.05、45.11±0.13、61.62±0.08、93.80±0.12)대세포생장적억제솔여대조조(2.03±0.07、2.78±0.08、3.11±0.01、5.67±0.04、12.23±0.04、18.65±0.04、24.45±0.04)비교명현승고(P균<0.05);단여순박비교,아신산기효쾌,재2～32 h억제솔명현고우동시간순박(P균<0.05).아신산화순박균가도치MG-63세포조망,조망솔조간비교차이유통계학의의(F=13.317,P<0.05);아신산재24、36、48 h(20.5±3.78、45.76±9.90、25.16±15.41),순박재24、36、48 h(12.55±1.51、18.85±3.40、12.37±5.43),MG-63세포조망솔(%)명현고우동시간대조조(6.57±1.48、8.03±2.08、6.54±1.30,P<0.05혹<0.01).아신산화순박균대MG-63세포주기G1、S、G2/M기유억제작용,억제솔조간비교차이유통계학의의(F치분별위54.579、43.429、21.795,P<0.05혹<0.01);대G1기억제솔(%)아신산(78.26±5.24)화순박(80.48±2.81)여대조조(57.49±6.65)비교명현승고(P<0.05혹<0.01).결론 아신산화순박도가억제골육류MG-63세포생장,유도세포조망;아신산작용쾌우순박,아신산화순박대MG-63세포적억제작용발생재세포주기중적G1기.
Objective To explore the inhibiting effects of arsenic trioxide and cisplatin on MG-63 cells. Methods Using MTT assay,flowcytometry,phase contrast microscopy and electron microscopy methods,the therapeutic effect of arsenic trioxide was studied for the osteosarcoma in the cultured MG-63 cells in vitro,and compared these effects with cisplatin. The inhibitory rotes of cell growth and the effect of apoptosis and cell cycle were compared between arsenic trioxide and cisplatin on MG-63 cells. Results The contrast phase microscope revealed the adhesion ability of normal groups was good and cellular morphology showed epithelium cells. But the celhdar morphology showed irregular arrangement in arsenic trioxide groups and cytoplasmic vacuoles in cisplatin group. Electron microscope revealed the globular plasmalemma ecphymas in cell surface of control groups,the enlarged crista mitochondriales and the double-deck membrane structure appeared clearly. But electron microscope revealed globular plasmalemma processes in cell surface of arsenic trioxide groups,thinned crista mitochondriales and clearly seen karyopycnosis and nuclear membrane of apoptotic cells. The globular plasmalemma processes in cell surface of cisplatin groups were separated,nuclear membrane thickened and chromatin were in sandy shape. Both arsenic trioxide and cisplatin inhibited effectively MG-63 cells growth. There was a significant difference in different groups of inhibition ratios to the growth of cells(all P < 0.05). In 2,4,8,16,32,64,128 hours,the inhibition ratios(%) of arsenic trioxide(56.31±0.03,70.00±0.06,79.84±0.03,87.31±0.13,84.70±0.09,90.68±0.06,91.18±0.05) and cisplatin groups(7.55±0.15,15.70±0.17,30.72±0.07,49.80±0.05,45.11± 0.13,61.62±0.08,93.80±0.12) were obviously increased as compared with those in the control group(2.03± 0.07,2.78±0.08,3.11±0.01,5.67±0.04,12.23±0.04,18.65±0.04,24.45±0.04,all P < 0.05). Moreover the inhibition ratio of arsenic trioxide group in 2 to 32 hour was significantly higher than that of cisplatin group and the effect was more faster(all P < 0.05). Both arsenic trioxide and cisplatin could induce apoptosis MG-63 cells. There was a significant difference in different groups of the inhibition ratio to the growth of cells(F = 13.317,P < 0.05). The inhibition ratios(%) of arsenic trioxide on 24,36,48 hour(20.50±3.78,45.76±9.90,25.16±15.41),and cisplatin groups on 24,36,48 hour(12.55±1.51,18.85±3.40,12.37±5.43),were obviously increased as compared with those in the control group at the same time(6.57±1.48,8.03±2.08,6.54±1.30,P< 0.05 or<0.01). Both arsenic trioxide and cisplatin inhibited MG-63 cells cycle. There was a significant difference in different groups of the inhibition ratio to the growth of cells(F = 54.579,43.429,21.795,P < 0.05 or < 0.01). And the total inhibition ratios(%) in G1 cycle of arsenic trioxide(78.26±5.24) and cisplatin groups(80.48±2.81) were obviously increased as compared with those in the control group(57.49±6.65,all P < 0.05 or < 0.01). Conclusions Arsenic trioxide and cisplatin can effectively inhibit the proliferation of MG-63 cell line and induce the apoptosis of MG-63 cell line. And the effects induced by arsenic trioxide group were faster than that of cisplatin groups. Moreover arsenic trioxide can arrest the cell cycle of MG-63 cell line at G1 phase.
