中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年
10期
1511-1515
,共5页
陈琪枫%方晓明%姚宁%方旭东%龚谋春
陳琪楓%方曉明%姚寧%方旭東%龔謀春
진기풍%방효명%요저%방욱동%공모춘
干细胞%骨髓干细胞%肝癌细胞%人骨髓间充质干细胞%生物学行为%侵袭能力%细胞增殖
榦細胞%骨髓榦細胞%肝癌細胞%人骨髓間充質榦細胞%生物學行為%侵襲能力%細胞增殖
간세포%골수간세포%간암세포%인골수간충질간세포%생물학행위%침습능력%세포증식
Bone Marrow%Mesenchymal Stem Cels%Carcinoma,Hepatocelular%Neoplasm Invasiveness
背景：肝癌的转移特性是影响预后的关键因素，观察人骨髓间充质干细胞对肝癌转移行为的影响，对于提高肝癌患者的生存期具有重要意义。目的：探讨人骨髓间充质干细胞对不同转移潜能肝癌细胞生物学特性的影响。方法：Transwel下室加入人骨髓间充质干细胞，上室加入高、低转移潜能肝癌细胞悬液，共培养36 h酶标仪检测吸光度值及细胞计数法观察肝癌细胞侵袭能力的变化。CCK-8法检测人骨髓间充质干细胞对高低转移潜能肝癌细胞增殖能力的影响。PCR检测人骨髓间充质干细胞与高低转移潜能肝癌细胞共培养前后转移相关因子骨桥蛋白、唾液酸蛋白、整合(α-V)以及增殖相关基因转化生长因子β1和程序化凋亡因子的表达。结果与结论：①共培养组的肝癌细胞迁移数量显著少于单独培养组，经酶标仪半定量检测肝癌细胞侵袭性显示，共培养组吸光度值明显低于单独培养组，差异有显著性意义(P <0.05)。②高转移潜能肝癌细胞与人骨髓间充质干细胞共培养组转移相关因子骨桥蛋白和骨唾液酸蛋白表达显著下降(P <0.05)，但整合(α-V)基因表达未出现明显改变(P >0.05)。低转移潜能肝癌细胞与人骨髓间充质干细胞共培养组转移相关因子骨桥蛋白、骨唾液酸蛋白以及整合(α-V)基因表达显著下降(P <0.05)。③经酶标仪半定量检测肝癌细胞增殖能力显示，共培养组吸光度值明显高于单独培养组，差异有显著性意义(P <0.05)。④高转移潜能肝癌细胞与人骨髓间充质干细胞共培养组的转化生长因子β1基因表达显著上调(P <0.05)；但程序化凋亡因子基因表达无明显改变(P >0.05)。低转移潜能肝癌细胞与人骨髓间充质干细胞共培养组的转化生长因子β1、程序化凋亡因子基因表达均显著上调(P <0.05)。以上结果提示人骨髓间充质干细胞与肝癌细胞共培养后，肝癌细胞侵袭能力明显下调，增殖能力明显提高。
揹景：肝癌的轉移特性是影響預後的關鍵因素，觀察人骨髓間充質榦細胞對肝癌轉移行為的影響，對于提高肝癌患者的生存期具有重要意義。目的：探討人骨髓間充質榦細胞對不同轉移潛能肝癌細胞生物學特性的影響。方法：Transwel下室加入人骨髓間充質榦細胞，上室加入高、低轉移潛能肝癌細胞懸液，共培養36 h酶標儀檢測吸光度值及細胞計數法觀察肝癌細胞侵襲能力的變化。CCK-8法檢測人骨髓間充質榦細胞對高低轉移潛能肝癌細胞增殖能力的影響。PCR檢測人骨髓間充質榦細胞與高低轉移潛能肝癌細胞共培養前後轉移相關因子骨橋蛋白、唾液痠蛋白、整閤(α-V)以及增殖相關基因轉化生長因子β1和程序化凋亡因子的錶達。結果與結論：①共培養組的肝癌細胞遷移數量顯著少于單獨培養組，經酶標儀半定量檢測肝癌細胞侵襲性顯示，共培養組吸光度值明顯低于單獨培養組，差異有顯著性意義(P <0.05)。②高轉移潛能肝癌細胞與人骨髓間充質榦細胞共培養組轉移相關因子骨橋蛋白和骨唾液痠蛋白錶達顯著下降(P <0.05)，但整閤(α-V)基因錶達未齣現明顯改變(P >0.05)。低轉移潛能肝癌細胞與人骨髓間充質榦細胞共培養組轉移相關因子骨橋蛋白、骨唾液痠蛋白以及整閤(α-V)基因錶達顯著下降(P <0.05)。③經酶標儀半定量檢測肝癌細胞增殖能力顯示，共培養組吸光度值明顯高于單獨培養組，差異有顯著性意義(P <0.05)。④高轉移潛能肝癌細胞與人骨髓間充質榦細胞共培養組的轉化生長因子β1基因錶達顯著上調(P <0.05)；但程序化凋亡因子基因錶達無明顯改變(P >0.05)。低轉移潛能肝癌細胞與人骨髓間充質榦細胞共培養組的轉化生長因子β1、程序化凋亡因子基因錶達均顯著上調(P <0.05)。以上結果提示人骨髓間充質榦細胞與肝癌細胞共培養後，肝癌細胞侵襲能力明顯下調，增殖能力明顯提高。
배경：간암적전이특성시영향예후적관건인소，관찰인골수간충질간세포대간암전이행위적영향，대우제고간암환자적생존기구유중요의의。목적：탐토인골수간충질간세포대불동전이잠능간암세포생물학특성적영향。방법：Transwel하실가입인골수간충질간세포，상실가입고、저전이잠능간암세포현액，공배양36 h매표의검측흡광도치급세포계수법관찰간암세포침습능력적변화。CCK-8법검측인골수간충질간세포대고저전이잠능간암세포증식능력적영향。PCR검측인골수간충질간세포여고저전이잠능간암세포공배양전후전이상관인자골교단백、타액산단백、정합(α-V)이급증식상관기인전화생장인자β1화정서화조망인자적표체。결과여결론：①공배양조적간암세포천이수량현저소우단독배양조，경매표의반정량검측간암세포침습성현시，공배양조흡광도치명현저우단독배양조，차이유현저성의의(P <0.05)。②고전이잠능간암세포여인골수간충질간세포공배양조전이상관인자골교단백화골타액산단백표체현저하강(P <0.05)，단정합(α-V)기인표체미출현명현개변(P >0.05)。저전이잠능간암세포여인골수간충질간세포공배양조전이상관인자골교단백、골타액산단백이급정합(α-V)기인표체현저하강(P <0.05)。③경매표의반정량검측간암세포증식능력현시，공배양조흡광도치명현고우단독배양조，차이유현저성의의(P <0.05)。④고전이잠능간암세포여인골수간충질간세포공배양조적전화생장인자β1기인표체현저상조(P <0.05)；단정서화조망인자기인표체무명현개변(P >0.05)。저전이잠능간암세포여인골수간충질간세포공배양조적전화생장인자β1、정서화조망인자기인표체균현저상조(P <0.05)。이상결과제시인골수간충질간세포여간암세포공배양후，간암세포침습능력명현하조，증식능력명현제고。
BACKGROUND:The metastatic potential of hepatocelular carcinoma cels is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cels on metastasis of hepatocelular carcinoma is of great significance for improving the lifetime of hepatocelular carcinoma patients. OBJECTIVE:To explore the biological effect of human bone marrow mesenchymal stem cels on hepatocelular carcinoma cels with different metastatic potentials. METHODS:Human bone marrow mesenchymal stem cels and hepatocelular carcinoma cel suspension with high and low metastatic potentials were respectively injected into the Transwel chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as wel as cel counting method was used to observe the changes in the invasion ability of hepatocelular carcinoma cels. The effects of human bone marrow mesenchymal stem cels on the proliferation of hepatocelular carcinoma cel suspension with high and low metastatic potentials were determined using cel counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cel death protein 5. RESULTS AND CONCLUSION:(1) The number of migrated hepatocelular carcinoma cels was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P> 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group (P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P< 0.05), but the expression of programmed cel death protein 5 showed no changes (P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cel death protein 5 was both increased dramaticaly (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cels reduce the invasion ability of hepatocelular carcinoma cels, and enhance their ability of proliferation.
