黃晶果萃取物對人類肝癌細胞生長之影響

尋找作物或食材做為開發保健產品及藥物之有效成分來源，為對於現今提升的健康意識之重要研究。文獻提到Pouteria caimito具多種生物活性，三萜類與類黃酮等化合物是該屬的主要活性成分。本研究是以黃晶果 (Pouteria caimito) 之植物部位 (種子、果皮、葉子與枝條) 分別以95%酒精及水進行萃取作為試驗樣品，探討其對於人類肝癌細胞株Hep3B生長的抑制作用。實驗以細胞存活率試驗篩選出種子酒萃物為黃晶果較有效抑制Hep3B細胞生長的樣品，以其進行後續研究之探討。經MTT assay結果顯示種子酒萃物能夠降低Hep3B細胞生存率，並且具有時間與劑量效應。此外，使用相位差顯微鏡觀察經種子酒萃物處理之Hep3B細胞的細胞形態，隨著樣品濃度提高，細胞數量隨之減少，並且產生變形及萎縮的現象。而經由流式細胞儀分析結果顯現出，種子酒萃物於細胞週期中可促進sub-G1 phase的增加，誘導Hep3B細胞產生細胞凋亡。藉由DAPI染色於螢光顯微鏡的觀察下，細胞核濃縮程度隨著黃晶果種子酒萃物處理濃度的增加而提升。並且經由caspase-3、-8及-9活性的測定之下，得知種子酒萃物經由caspase-3及caspase-9的路徑活化細胞凋亡。綜合上述，黃晶果種子酒萃物可透過細胞凋亡有效抑制人類肝癌細胞株Hep3B之生長，在體外實驗中表現出良好的抑癌效果，後續有望持續研究，將黃晶果作為開發相關保健產品或藥物的有效潛力作物。

關鍵字

人類肝癌細胞株Hep3B ；黃晶果種子；細胞凋亡

並列摘要

Looking for crops or food ingredients as sources of active ingredients for the development of health care products and medicines is an important research for raised health awareness today. The literature mentioned that Pouteria caimito fruit has a variety of biological activities, and triterpenoids and flavonoids are the main active components of this genus. In this study, the plant parts (seeds, peels, leaves and branches) of Pouteria Caimito were extracted with 95% alcohol and water respectively as test samples to explore their inhibitory effect on the growth of human hepatoma cancer cell line Hep3B. The experiment screened out the sample with effective inhibition by cell viability test, and alcoholic extract of Pouteria Caimito seed showed more effective inhibitory effect on the growth of Hep3B cells, which will be used in the follow-up experiments. The results of MTT assay showed that the alcoholic extract of seed reduced the cell viability of Hep3B cells, and had time- and dose-dependent effects. In addition, Hep3B cells treated with alcoholic extract of seed were observed by phase contrast microscope. With the increase of sample concentration, the number of cells decreased, and deformation and atrophy occurred. The results of flow cytometry analysis showed that the alcoholic extract of seed promoted the increase of sub-G1 phase in the cell cycle and induced apoptosis of Hep3B cells. The degree of nuclear condensation of Hep3B cells was increased with the enhanced treatment concentration of alcoholic extract of Pouteria Caimito seed by DAPI staining under fluorescence microscope. Through the determination of the activities of caspase-3, -8 and -9, it is known that the alcoholic extract of seed activates apoptosis through the pathway of caspase-3 and caspase-9. In summary, alcoholic extract of Pouteria Caimito seed can effectively inhibit the growth of human hepatoma cancer cell line Hep3B through apoptosis, and showed a great effect in in vitro experiments. Pouteria Caimito is expected to continue to be studied as an effective potential crop for the development of related health care products or drugs in the future.