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)
2013年
21期
9590-9594
,共5页
张鹏%李书忠%张金锋%曲邵政%高甲科%张志
張鵬%李書忠%張金鋒%麯邵政%高甲科%張誌
장붕%리서충%장금봉%곡소정%고갑과%장지
β连环素%WIF-1%人骨肉瘤MG-63
β連環素%WIF-1%人骨肉瘤MG-63
β련배소%WIF-1%인골육류MG-63
beta Catenin%WIF-1%Human osteosarcoma MG-63 cells
目的： WIF-1是Wnt信号通路中最常见抑制因子之一，本实验研究WIF-1对人骨肉瘤MG-63细胞Wnt信号通路中β-catenin表达量的影响。方法细胞培养人骨肉瘤MG-63细胞，0 ng/ml浓度的WIF-1作为空白对照组，分别以0、48、60、80、120 ng/ml浓度的WIF-1刺激MG-63细胞，分别用FQ-PCR及Western blot法从基因水平、蛋白水平测定细胞中β-catenin的表达量；细胞培养人骨肉瘤MG-63细胞，用WIF-1刺激细胞，分别在第0、12、24、48、72、96 h对各组细胞进行计数，观察空白组及WIF-1刺激组细胞数目的变化情况。结果不同浓度（0、48、60、80、120 ng/ml）WIF-1刺激下，FQ-PCR法显示β-catenin mRNA的相对表达总量2-ΔΔCT分别为1、0.55±0.30、0.44±0.24、0.35±0.15、0.24±0.21。与空白组相比较，WIF-1刺激下β-catenin mRNA表达量均降低（P＜0.05）。随着WIF-1浓度的增加，β-catenin mRNA表达量下降，但组间的抑制作用无明显差别（P＞0.05）；Western blot法显示β-catenin/β-actin蛋白相对表达量分别为1.40±0.11、0.86±0.10、0.51±0.11、0.45±0.06、0.18±0.03；与空白组相比较，差异有统计学意义（P＜0.05）；空白组及WIF-1刺激组细胞数目有差异，空白组细胞增殖情况优于WIF-1刺激组。结论 WIF-1可降低人骨肉瘤MG-63细胞Wnt信号通路中β-catenin表达量，并抑制人骨瘤MG-63细胞的增殖。
目的： WIF-1是Wnt信號通路中最常見抑製因子之一，本實驗研究WIF-1對人骨肉瘤MG-63細胞Wnt信號通路中β-catenin錶達量的影響。方法細胞培養人骨肉瘤MG-63細胞，0 ng/ml濃度的WIF-1作為空白對照組，分彆以0、48、60、80、120 ng/ml濃度的WIF-1刺激MG-63細胞，分彆用FQ-PCR及Western blot法從基因水平、蛋白水平測定細胞中β-catenin的錶達量；細胞培養人骨肉瘤MG-63細胞，用WIF-1刺激細胞，分彆在第0、12、24、48、72、96 h對各組細胞進行計數，觀察空白組及WIF-1刺激組細胞數目的變化情況。結果不同濃度（0、48、60、80、120 ng/ml）WIF-1刺激下，FQ-PCR法顯示β-catenin mRNA的相對錶達總量2-ΔΔCT分彆為1、0.55±0.30、0.44±0.24、0.35±0.15、0.24±0.21。與空白組相比較，WIF-1刺激下β-catenin mRNA錶達量均降低（P＜0.05）。隨著WIF-1濃度的增加，β-catenin mRNA錶達量下降，但組間的抑製作用無明顯差彆（P＞0.05）；Western blot法顯示β-catenin/β-actin蛋白相對錶達量分彆為1.40±0.11、0.86±0.10、0.51±0.11、0.45±0.06、0.18±0.03；與空白組相比較，差異有統計學意義（P＜0.05）；空白組及WIF-1刺激組細胞數目有差異，空白組細胞增殖情況優于WIF-1刺激組。結論 WIF-1可降低人骨肉瘤MG-63細胞Wnt信號通路中β-catenin錶達量，併抑製人骨瘤MG-63細胞的增殖。
목적： WIF-1시Wnt신호통로중최상견억제인자지일，본실험연구WIF-1대인골육류MG-63세포Wnt신호통로중β-catenin표체량적영향。방법세포배양인골육류MG-63세포，0 ng/ml농도적WIF-1작위공백대조조，분별이0、48、60、80、120 ng/ml농도적WIF-1자격MG-63세포，분별용FQ-PCR급Western blot법종기인수평、단백수평측정세포중β-catenin적표체량；세포배양인골육류MG-63세포，용WIF-1자격세포，분별재제0、12、24、48、72、96 h대각조세포진행계수，관찰공백조급WIF-1자격조세포수목적변화정황。결과불동농도（0、48、60、80、120 ng/ml）WIF-1자격하，FQ-PCR법현시β-catenin mRNA적상대표체총량2-ΔΔCT분별위1、0.55±0.30、0.44±0.24、0.35±0.15、0.24±0.21。여공백조상비교，WIF-1자격하β-catenin mRNA표체량균강저（P＜0.05）。수착WIF-1농도적증가，β-catenin mRNA표체량하강，단조간적억제작용무명현차별（P＞0.05）；Western blot법현시β-catenin/β-actin단백상대표체량분별위1.40±0.11、0.86±0.10、0.51±0.11、0.45±0.06、0.18±0.03；여공백조상비교，차이유통계학의의（P＜0.05）；공백조급WIF-1자격조세포수목유차이，공백조세포증식정황우우WIF-1자격조。결론 WIF-1가강저인골육류MG-63세포Wnt신호통로중β-catenin표체량，병억제인골류MG-63세포적증식。
ObjectiveWIF-1(Wnt inhibitory factor-1) was one of the most common antagonists of wnt signal pathway, this experiment was to study the expression of β-catenin interfered with WIF-1 in human osteosarcoma MG-63 cells.MethodsHuman MG-63 cells interfered with WIF-1 in the concentration of 0 ng/ml,48 ng/ml,60 ng/ml,80 ng/ml,120 ng/ml was cultured, the concentration of 0 ng/ml was considered as the blank group. The expression of β-catenin was retrospectively detected by the methods of real-time fluorescent quantitative polymerase chain reaction(FQ-PCR) and Western blot at the genetic and protein level. Culturing human MG-63 cells, interfered with the same concentration of WIF-1, the quantity of cells was partly noted in the hour of 0, 12, 24, 48, 72, 96 h. The number was compared between in the blank group and experiment group. Results Interfered with different concentrations(0, 48, 60, 80, 120 ng/ml) of WIF-1. It was showed by FQ-PCR that the relative fold-chages ofβ-catenin mRNA was retrospectively recorded as 1, 0.55±0.30, 0.44±0.24, 0.35±0.15, 0.24±0.21. Compared with the blank group, the stimulating groups were all found to have a lower expression(P<0.05). With the improvement of concentrations of WIF-1, the expressions of β-catenin presents a decreasing trend .However, these was no significant differences in the stimulating groups themselves; Western blot showed us that the relative fold-chages ofβ-catenin/β-actin was separately memorized as 1.40±0.11, 0.86±0.10, 0.51±0.11, 0.45±0.06, 0.18±0.03; compared with the blank group, differences were statistically significant(P<0.05). Compared with the stimulating group, the blank group was found to have a higher proliferation. ConclusionWIF-1 could decrease the expressions of β-catenin in the canonical wnt signal pathway of human osteosarcoma MG-63 cells and inhibit the proliferation of human osteosarcoma MG-63 cell.
