根據行政院衛生署統計,肝癌為男性十大癌症死因之首,女性則居第二位,且死亡率逐年增加。桑黃Phellinus Linteus為一種寄生於桑樹上的多年生蕈菇類,具有保肝及體內治療肝癌的效果。而Hispidin為傳統藥用蕈菇類桑黃之活性成分之ㄧ。但Hispidin對於人類肝癌細胞株是否具有抑制的作用並無相關文獻報導且對於抑制癌症的機制仍不清楚,因此在本實驗中以Hispidin作用於人類肝癌細胞株HepG2來評估其生長抑制的效應及探討Hispidin誘導HepG2細胞之凋亡作用及其可能機制。由MTT分析結果顯示Hispidin對HepG2有明顯的抑制生長作用。而且Hispidin影響HepG2的細胞週期休止於S 期。經由DAPI染色、DNA Laddering試驗及Annexin V / PI的雙染分析都可觀察到Hispidin對於HepG2具有促進細胞凋亡效果。而且經實驗證實Hispidin會使HepG2細胞內ROS生成量增加。在Hispidin誘導肝癌細胞凋亡路徑方面,可能有活化PI3K/AKt途徑、MAPK途徑、粒線體途徑以及Caspase途徑。此外,由實驗結果得知Hispidin 亦可以抑制HepG2細胞MMP-2和MMP-9的活性,所以推測Hispidin可能具有抑制轉移的能力。
According to the report of the Department of Health (DOH) of Taiwan, the most common death causes are cancers. Hepatocellular carcinoma (HCC) is the first and commonest cancer in men and the second and commonest cancer in women. Hispidin is a component from Phellinus Linteus, one of the traditional Chinese medical herbs. However, the underlying mechanism of the anti-tumor effects of Hispidin remains to be clarified. In the present study, Hispidin was used to evaluate growth inhibition on human liver cancer cell line HepG2. The inhibitory effects of Hispidin on the growth of tumor cells were determined by a MTT assay. Hispidin induced cell cycle arrest in the S phase. In the DAPI staining、DNA fragmentation and Annexin V/ PI double staining, it was identified that Hispidin could induce HepG2 apoptosis. Hispidin could induce HepG2 apoptosis through mediating ROS generation. By Western blot analysis, as a result that the HepG2 cells underwent apoptosis via activating PI3K/AKt pathway、MAPK pathway、mitochondria pathway and caspase pathway. Otherwise, according to the result that Hispidin might inhibit MMP-2 and MMP-9 protein expression in HepG2 cells. So Hispidin might be able to inhibit migration.