刺参酸性黏多糖对人肝癌HepG2细胞增殖的影响

EFFECT OF STICHOPUS JAPONICUS ACID MUCOPOLYSACCHARIDE ON PROLIFERATION OF LIVER CANCER CELL HepG2

目的了解刺参酸性黏多糖(SJAMP)对体外培养的人肝癌HepG2细胞增殖的影响。方法体外培养HepG2细胞,MTT法检测SJAMP对细胞增殖的影响,流式细胞术检测细胞周期,免疫细胞化学法检测细胞增殖细胞核抗原(PCNA)表达,以正常肝细胞(张氏细胞)作为对照。结果 SJAMP能抑制HepG2细胞增殖,且随着干预剂量(0、0.5、2.0、8.0mg/L)和干预时间(12、24、48h)延长其抑制作用增强(F=68.430～596.680,q=3.714～55.029,P<0.05);不同浓度SJAMP作用对张氏细胞增殖无明显的抑制作用(P<0.05)。随着SJAMP剂量的增加,HepG2细胞停留在G1期比例增加,S期、G2期的细胞减少(F=4.483～18.791,q=1.921～9.876,P<0.05);对于张氏细胞,SJAMP作用后其细胞周期未发生显著改变(P>0.05)。SJAMP作用后HepG2细胞的PCNA表达降低,且随着SJAMP作用剂量的增加而降低(F=2 688.640,q=55.365～156.296,P<0.05);张氏细胞PC-NA表达未发生显著变化(P>0.05)。结论 SJAMP通过诱导HepG2细胞发生细胞周期G1期阻滞并抑制其PCNA表达,从而抑制HepG2细胞增殖,发挥抗肿瘤作用。

Objective To study the effect of stichopus japonicus acid mucopolysaccharide （SJAMP） on proliferation of human liver cancer cell HepG2. Methods HepG2 cells were cultured in vitro. MTT assay was used to observe the effect of SJAMP on proliferation of HepG2 cells. The cell cycle was measured using flow cytometry （FCM）. The expression of PCNA was tested by immunocytochemistry. Normal liver cells （Zhang＇s liver cells） were served as the control. Results SJAMP could inhibit the proliferation of HepG2 cells, its depressant effect reinforced along with the increase of intervention dose （0,0.5,2.0, 8.0 mg/L） and extension of intervention time （12,24,48 b） （F=68.430--596.680;q=3.714-55.029;P〈0.05）; Different concentrations of SJAMP did not show apparent inhibition on proliferation of Zhang＇s cells （P〈0.05）. The results of cell cycle detection indicated that with the increase of SJAMP dose, the HepG2 ceils that trapped in G1 phase increased （F=4.483-18.791 ;q= 1.921-9.876;P〈0.05） ; as for Zhang＇s cells, no changes of cell cycle were noted after interfered with SJAMP （P〈0.05） Immunocytochemical technique indicated that after SJAMP intervention, the expression of PCNA in HepG2 cells declined, the expression decreased as the dose of SJ AMP increased （F = 2 688.640, q = 55.365 - 156.296, P 〈0.05 ）, no notable effect on the expression of Zhang＇s cells was observed （P〉0.05）. Conclusion SJAMP plays an anti-tumor action through inhibiting the proliferation,blocking cell cycle and decreasing expression of PCNA of HepG2 ceils.