中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
10期
2057-2059
,共3页
熊金蓉%胡祥%喻爱喜%刘胜军%吴刚
熊金蓉%鬍祥%喻愛喜%劉勝軍%吳剛
웅금용%호상%유애희%류성군%오강
骨肉瘤%抑癌基因%增殖%侵袭
骨肉瘤%抑癌基因%增殖%侵襲
골육류%억암기인%증식%침습
Osteosarcoma cells%Suppressor gene%Proliferation%Invasion
目的 观察抑癌基因磷酸酶及张力蛋白同源的基因(PTEN)过表达对骨肉瘤细胞MG63和U2OS增殖和侵袭的影响.方法 培养人骨肉瘤MG63和U2OS细胞,构建PcDNA3.0-PTEN表达系统,并导入骨肉瘤细胞中,采用免疫组织化学和Western blot法检测PTEN的表达；用噻唑蓝(MTT)比色法及侵袭小室实验检测pcDNA3.0-PTEN转染细胞的增殖活性和侵袭力.结果 与对照组比较,pcDNA3.0-PTEN的稳定转染明显能使骨肉瘤细胞MG63和U2OS的PTEN过表达.增殖实验表明PTEN转染组细胞增殖活性低于对照组,培养第6天MG63吸光度(A)值分别为5.290 ±0.032比6.690 ±0.051(P＜0.05),U2OS分别为4.790±0.032比5.690 ±0.051(P＜0.05)；侵袭实验证实转染组比对照组穿膜能力显著下降,穿膜细胞数:MG63:0.560±0.023比1.020±0.018(P ＜0.05)；U2OS:0.690 ±0.021比0.980 ±0.016(P ＜0.05).结论 PTEN过表达能显著抑制骨肉瘤细胞的增殖和侵袭.
目的 觀察抑癌基因燐痠酶及張力蛋白同源的基因(PTEN)過錶達對骨肉瘤細胞MG63和U2OS增殖和侵襲的影響.方法 培養人骨肉瘤MG63和U2OS細胞,構建PcDNA3.0-PTEN錶達繫統,併導入骨肉瘤細胞中,採用免疫組織化學和Western blot法檢測PTEN的錶達；用噻唑藍(MTT)比色法及侵襲小室實驗檢測pcDNA3.0-PTEN轉染細胞的增殖活性和侵襲力.結果 與對照組比較,pcDNA3.0-PTEN的穩定轉染明顯能使骨肉瘤細胞MG63和U2OS的PTEN過錶達.增殖實驗錶明PTEN轉染組細胞增殖活性低于對照組,培養第6天MG63吸光度(A)值分彆為5.290 ±0.032比6.690 ±0.051(P＜0.05),U2OS分彆為4.790±0.032比5.690 ±0.051(P＜0.05)；侵襲實驗證實轉染組比對照組穿膜能力顯著下降,穿膜細胞數:MG63:0.560±0.023比1.020±0.018(P ＜0.05)；U2OS:0.690 ±0.021比0.980 ±0.016(P ＜0.05).結論 PTEN過錶達能顯著抑製骨肉瘤細胞的增殖和侵襲.
목적 관찰억암기인린산매급장력단백동원적기인(PTEN)과표체대골육류세포MG63화U2OS증식화침습적영향.방법 배양인골육류MG63화U2OS세포,구건PcDNA3.0-PTEN표체계통,병도입골육류세포중,채용면역조직화학화Western blot법검측PTEN적표체；용새서람(MTT)비색법급침습소실실험검측pcDNA3.0-PTEN전염세포적증식활성화침습력.결과 여대조조비교,pcDNA3.0-PTEN적은정전염명현능사골육류세포MG63화U2OS적PTEN과표체.증식실험표명PTEN전염조세포증식활성저우대조조,배양제6천MG63흡광도(A)치분별위5.290 ±0.032비6.690 ±0.051(P＜0.05),U2OS분별위4.790±0.032비5.690 ±0.051(P＜0.05)；침습실험증실전염조비대조조천막능력현저하강,천막세포수:MG63:0.560±0.023비1.020±0.018(P ＜0.05)；U2OS:0.690 ±0.021비0.980 ±0.016(P ＜0.05).결론 PTEN과표체능현저억제골육류세포적증식화침습.
Objective To observe the effect of up-regulation of the suppressor gene phosphatase and tensin homolog deleted on chromosome ten (PTEN) on the proliferation and invasion of osteosarcoma cell lines MG63 and U2OS.Methods Human osteosarcoma cell lines MG63 and U2OS were established,and then pcDNA3.0-PTEN was constructed and transfected into these cells.The effects of pcDNA3.0-PTEN on the proliferation and invasion in osteosarcoma cells were examined by methyl thiazol tetrazolium (MTT) assay and invasion assay following the PTEN overexpression was confirmed by immunocytochemistry and Western blotting.Results The overexpression of PTEN in MG63 and U20S cells was confirmed by Western blotting and immunocytochemistry.Compared with control group,the A value in MG63 cells transfected with PTEN after culture for 6 days examined by using MTT was (5.290 ± 0.032) vs.(6.690 ±0.051) in control group (P ＜ 0.05),and that in U2OS cells was (4.790 ± 0.032) vs.(5.690 ± 0.051)in control group (P ＜ 0.05).The number of penetrating cells in the MG63 group and U2OS group was (0.560±0.023) vs.(1.020 ±0.018) in control group (P＜0.05),and (0.690 ±0.021) vs.(0.980 ± 0.016) in control group,respectively (P ＜ 0.05).Conclusion Up-regulated PTEN suppresses the proliferation and invasion of osteosarcoma cells.
