中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
3期
480-482
,共3页
王凯%李玉明%张明凯%姜吉祥%苏庆亮
王凱%李玉明%張明凱%薑吉祥%囌慶亮
왕개%리옥명%장명개%강길상%소경량
胃癌%间充质干细胞%增殖细胞核抗原%基质金属蛋白酶
胃癌%間充質榦細胞%增殖細胞覈抗原%基質金屬蛋白酶
위암%간충질간세포%증식세포핵항원%기질금속단백매
Gastric cancer%Mesenchymal stem cells%Proliferating cell nuclear antigen%Matrix metalloproteinases
目的 观察人胃癌组织中分离培养的间充质干细胞(hGC-MSCs)对胃癌细胞株SGC-7901增殖与侵袭能力的影响.方法 提取hGC-MSCs条件培养液(MSC-CM),用不同浓度(20％、40％、60％)的MSC-CM作用于SGC-7901细胞,噻唑蓝(MTT)法及Transwell侵袭实验检测各组细胞增殖与侵袭能力;实时聚合酶链反应(Real-time PCR)、Western blot和细胞免疫化学法检测增殖细胞核抗原(PCNA)和基质金属蛋白酶(MMP)-9在各组细胞中的表达.结果 MTT法证实MSC-CM对SGC-7901细胞增殖有抑制作用(P＜0.05),抑制率与MSC-CM的浓度和作用时间呈正比.Transwell侵袭实验表明,经MSC-CM处理后,对照组及20％、40％、60％ MSC-CM组穿膜细胞数分别为(75.71±7.32)、(63.68 ±7.12)、(54.29 ±6.97)和(41.36±3.29)个,随MSC-CM浓度升高,细胞侵袭能力明显下降(P＜0.05).Real-time PCR结果证实MSC-CM能明显下调PCNA和MMP-9的表达(P＜0.01).Western blot检测20％、40％、60％ MSC-CM组PCNA表达分别为对照组的(73.99 ±16.71)％、(51.56±12.18)％和(40.11±9.37)％,MMP-9表达分别为对照组的(74.35±19.12)％、(53.59 ±10.41)％和(29.58±7.93)％,细胞免疫化学结果显示PCNA在对照组、20％、40％、60％ MSC-CM组中的平均吸光度值分别为0.201 ±0.049、0.185 ±0.031、0.154±0.019和0.128 ±0.012,MMP-9在4组中的平均吸光度值分别为0.182±0.032、0.164±0.028、0.136±0.017和0.098±0.008,各组之间差异有统计学意义(P＜0.05).结论 HGC-MSCs能抑制SGC-7901细胞的增殖及侵袭能力,其机制可能与下调PCNA和MMP-9的表达有关.
目的 觀察人胃癌組織中分離培養的間充質榦細胞(hGC-MSCs)對胃癌細胞株SGC-7901增殖與侵襲能力的影響.方法 提取hGC-MSCs條件培養液(MSC-CM),用不同濃度(20％、40％、60％)的MSC-CM作用于SGC-7901細胞,噻唑藍(MTT)法及Transwell侵襲實驗檢測各組細胞增殖與侵襲能力;實時聚閤酶鏈反應(Real-time PCR)、Western blot和細胞免疫化學法檢測增殖細胞覈抗原(PCNA)和基質金屬蛋白酶(MMP)-9在各組細胞中的錶達.結果 MTT法證實MSC-CM對SGC-7901細胞增殖有抑製作用(P＜0.05),抑製率與MSC-CM的濃度和作用時間呈正比.Transwell侵襲實驗錶明,經MSC-CM處理後,對照組及20％、40％、60％ MSC-CM組穿膜細胞數分彆為(75.71±7.32)、(63.68 ±7.12)、(54.29 ±6.97)和(41.36±3.29)箇,隨MSC-CM濃度升高,細胞侵襲能力明顯下降(P＜0.05).Real-time PCR結果證實MSC-CM能明顯下調PCNA和MMP-9的錶達(P＜0.01).Western blot檢測20％、40％、60％ MSC-CM組PCNA錶達分彆為對照組的(73.99 ±16.71)％、(51.56±12.18)％和(40.11±9.37)％,MMP-9錶達分彆為對照組的(74.35±19.12)％、(53.59 ±10.41)％和(29.58±7.93)％,細胞免疫化學結果顯示PCNA在對照組、20％、40％、60％ MSC-CM組中的平均吸光度值分彆為0.201 ±0.049、0.185 ±0.031、0.154±0.019和0.128 ±0.012,MMP-9在4組中的平均吸光度值分彆為0.182±0.032、0.164±0.028、0.136±0.017和0.098±0.008,各組之間差異有統計學意義(P＜0.05).結論 HGC-MSCs能抑製SGC-7901細胞的增殖及侵襲能力,其機製可能與下調PCNA和MMP-9的錶達有關.
목적 관찰인위암조직중분리배양적간충질간세포(hGC-MSCs)대위암세포주SGC-7901증식여침습능력적영향.방법 제취hGC-MSCs조건배양액(MSC-CM),용불동농도(20％、40％、60％)적MSC-CM작용우SGC-7901세포,새서람(MTT)법급Transwell침습실험검측각조세포증식여침습능력;실시취합매련반응(Real-time PCR)、Western blot화세포면역화학법검측증식세포핵항원(PCNA)화기질금속단백매(MMP)-9재각조세포중적표체.결과 MTT법증실MSC-CM대SGC-7901세포증식유억제작용(P＜0.05),억제솔여MSC-CM적농도화작용시간정정비.Transwell침습실험표명,경MSC-CM처리후,대조조급20％、40％、60％ MSC-CM조천막세포수분별위(75.71±7.32)、(63.68 ±7.12)、(54.29 ±6.97)화(41.36±3.29)개,수MSC-CM농도승고,세포침습능력명현하강(P＜0.05).Real-time PCR결과증실MSC-CM능명현하조PCNA화MMP-9적표체(P＜0.01).Western blot검측20％、40％、60％ MSC-CM조PCNA표체분별위대조조적(73.99 ±16.71)％、(51.56±12.18)％화(40.11±9.37)％,MMP-9표체분별위대조조적(74.35±19.12)％、(53.59 ±10.41)％화(29.58±7.93)％,세포면역화학결과현시PCNA재대조조、20％、40％、60％ MSC-CM조중적평균흡광도치분별위0.201 ±0.049、0.185 ±0.031、0.154±0.019화0.128 ±0.012,MMP-9재4조중적평균흡광도치분별위0.182±0.032、0.164±0.028、0.136±0.017화0.098±0.008,각조지간차이유통계학의의(P＜0.05).결론 HGC-MSCs능억제SGC-7901세포적증식급침습능력,기궤제가능여하조PCNA화MMP-9적표체유관.
Objective To investigate effects of the mesenchymal stem cells isolated and cultured from human gastric cancer (hGC-MSCs) on proliferation and invasion of human gastric cancer cell strain SGC-7901.Methods The condition medium of hGC-MSCs (MSC-CM) was isolated,MSC-CM was applied on SGC-7901 cells at different concentrations (20％,40％ and 60％),the changes of proliferation and invasion were tested by methylthiazol tetrazolium (MTT) and Transwell chambers respectively; the expressions of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinases 9 (MMP-9) were detected by Real-time polymerase chain reaction (PCR),Western blotting and Immunocytochemistry.Results MTT results showed that the proliferation of SGC-7901 was significantly inhibited by MSC-CM and the inhibition rate was positively correlated with MSC-CM concentration and reaction time (P ＜ 0.05).The transwell chambers results revealed that cell invasion was significantly decreased after cells were dealed with the MSC-CM,penetrating cells in the control group and the MSC-CM groups at different concentrations (20％,40％,60％) were 75.71 ± 7.32,63.68 ± 7.12,54.29 ± 6.97 and 41.36 ± 3.29 (P ＜ 0.05),respectively.Real-time PCR results showed that MSC-CM could down-regulate the expression of PCNA and MMP-9 (P ＜ 0.01).Western blot revealed the expression of PCNA in MSC-CM groups at different concentrations (20％,40％,60％) were down-regulated to (73.99 ± 16.71) ％,(51.56 ± 12.18) ％ and (40.11 ±9.37)％,MMP-9 (74.35 ±19.12)％,(3.59 ±10.41)％ and (29.58 ±7.93)％,Immunocytochemistry revealed the average absorbance of PCNA in MSC-CM groups at different concentrations (20％,40％,60％) were 0.201 ±0.049,0.185 ± 0.031,0.154 ± 0.019 and 0.128 ± 0.012,MMP-9 were 0.182 ± 0.032,0.164 ± 0.028,0.136 ± 0.017 and 0.098 ± 0.008 (all P ＜ 0.05).Conclusion Proliferation and invasion of SGC-7901 were inhibited by hGC-MSCs and the mechanism may be related to the reduction of PCNA and MMP-9 expression.
