本研究以愛玉子多胜肽區分(polypeptide fraction, PF)於體外細胞模式(in vitro)下探討PF抑制白血病細胞(U937)生長並促使其分化及誘導細胞凋亡之能力,並以動物模式(in vivo)探討愛玉子PF於生物體內經由免疫調節方式達到抗CT26腫瘤的效果。將濃度為100 μg/mL之愛玉子PF刺激人類單核球細胞一天所製備之單核細胞條件培養液(mononuclear cell conditioned media, MNC-CMs),可有效抑制U937細胞生長(94%),且以NBT還原陽性率測試、NSE陽性率、型態學觀察及細胞表面抗原檢測下,皆可顯示U937細胞株可被誘導分化為正常的單核或巨噬細胞。而此單核細胞條件培養液中亦可測得高量的IL-1β(4446.5±58.5 pg/mL)、TNF-α(1324.8±90.6 pg/mL)、GM-CSF(305.9±13.2 pg/m)及IFN-γ(1557.7±16.2 pg/mL)等細胞激素。將愛玉子PF(100 μg/mL)與U937共同培養,其生長抑制率可達90%以上,且會使U937細胞週期停滯,而於DNA片段化、Annexin-V檢測可知PF可誘使細胞凋亡之發生,具有濃度及時間效應;而由流式細胞儀檢測可知,PF亦會影響粒腺體膜電位且活化caspase-3。故可得知,由PF所製備之單核細胞條件培養液可誘導白血病細胞株分化,而將PF與白血病細胞株共同培養則可誘導細胞凋亡,兩者皆可抑制白血病細胞生長。 另外為了研究愛玉子PF於生物體內經由免疫調節方式達到抗腫瘤的效果,在Balb/c小鼠皮下移植大腸癌細胞(CT26)形成腫瘤,分別於施打腫瘤同時(處理組)或待腫瘤長出後(治療組)及腫瘤移植前(預防組)餵食愛玉子PF。結果發現,處理組之小鼠餵食愛玉子PF劑量為150 mg/kg bw/day時能明顯抑制腫瘤生長(35.9%),同時能促進移植腫瘤小鼠之脾臟細胞增生及提高其脾臟細胞內TH、TC及巨噬細胞比例。而治療組當投予小鼠每公斤體重100 mg劑量之愛玉子PF可有效抑制腫瘤生長,但其促進脾臟細胞增生能力則不明顯。而預防組之結果顯示,當小鼠先給予劑量為100及200 mg/kg bw/day之愛玉子PF後,其腫瘤抑制率可高達58.1及62.8%,若配合5-FU之使用,抑制率更高達81%左右,但愛玉子PF無法改善因5-FU所引起的脾臟腫大及脾臟增生能力降低之現象。顯示愛玉子PF於細胞模式及動物模式下皆具有抑制腫瘤生長及具有免疫調節能力。
Polypeptide fraction (PF) from jelly-fig (Ficus awkeotsang Makino) achenes was prepared to determine its effect on proliferation, differentiation, and apoptosis of human leukemic cells (U937) in vitro and immunomodulation and antitumor of CT26 in vivo. First, human peripheral blood mononuclear cells were stimulated by PF (100 μg/mL) for 1 day to prepared conditioned media (PF-MNC-CM-1). With which high growth inhibition (94%) on U937 cells and nitroblue tetrazolium (NBT) positive percentage, nonspecific esterase (NSE) positive percentage and surface antigen expressions of differentiated cells were observed. By morphological examination, U937 cells were confirmed to be differentiated into mature monocytes or/and macrophages. Through an ELISA, we found that MNC-CM-1 (stimulated by 100 μg/mL PF) contained high levels of IL-1β (4446.5±58.5 pg/mL), TNF-