探討天然粗萃物對於黑色素瘤之抑癌作用

黑色素瘤 (melanoma) 是一種侵襲性極高的皮膚癌，早期發現可以手術切除，一旦發生轉移治療將變得困難。目前的臨床藥物常面臨嚴重副作用和抗藥性問題，因此須積極尋找天然、毒性低且能提升臨床藥物效果並降低抗藥性發生的輔助療法。過去研究發現天然粗萃物有效抑制多種癌症，但仍缺乏對黑色素瘤的研究，因此本論文旨在探討天然粗萃物對黑色素瘤的抑癌作用。首先利用MTT assay測試天然粗萃物對B16/F10細胞的細胞毒性，並透過Colony formation assay發現其有效抑制B16/F10細胞形成群集。雙向協同實驗和In vitro resistance assay中，天然粗萃物與臨床藥物5-FU合併使用對黑色素瘤細胞治療有協同效果並降低B16/F10細胞對5-FU產生抗藥性。此外，Annexin V stain和TUNEL assay證實天然粗萃物誘導B16/F10細胞凋亡，Western blot assay則顯示其調控細胞增殖、凋亡相關的蛋白表現。在腫瘤轉移方面，Wound healing assay及Transwell assay確定天然粗萃物抑制B16/F10細胞移行和侵襲，Western blot assay和RT-PCR近一步證實其調控轉移相關基因和蛋白的表達。在動物實驗中，天然粗萃物有效抑制黑色素瘤在老鼠體內生長並提高存活天數，H&E stain顯示其對腫瘤細胞有高度選擇性，IHC stain則確認天然粗萃物調控細胞增殖、凋亡、轉移相關蛋白的表達。綜合以上結果，天然粗萃物具有抑制黑色素瘤細胞增殖、促凋亡及抑制轉移的效果，對黑色素瘤治療有巨大潛力。

Melanoma is a highly aggressive form of skin cancer. In the early stages of the disease, it is typically treated with surgical resection. However, once metastasis occurs, treatment becomes challenging. Current clinical drugs often face severe side effects and drug resistance issues. Therefore, it is important to find natural, low-toxicity complementary therapies that can enhance the treatment effects of clinical drugs and reduce drug resistance. Previous studies have found that natural extracts can effectively inhibit the progression of various cancers, but research on melanoma is still lacking. This paper aims to investigate the anticancer effects of natural extracts on melanoma. First, the cytotoxic effect of natural extracts on B16/F10 cells was tested using the MTT assay. The colony formation assay showed that these extracts effectively inhibit the colony formation of B16/F10 cells. In the dual synergy assay and in vitro resistance assay, natural extracts combined with the clinical drug 5-FU had a synergistic effect on melanoma cells and reduced the resistance of B16/F10 cells to 5-FU. Additionally, the Annexin V stain and TUNEL assay confirmed that natural extracts induce apoptosis in B16/F10 cells. The Western blot assay showed that these extracts regulate the expression of proteins related to cell proliferation and apoptosis. As for tumor metastasis, the wound healing assay and transwell assay confirmed that natural extracts inhibit the migration and invasion abilities of B16/F10 cells. Western blot and RT-PCR further confirmed that these extracts regulate the expression of genes and proteins related to metastasis. In in vivo experiments, natural extracts effectively inhibited the tumor growth of melanoma in mice and increased their survival rate. The H&E staining showed a high selectivity of natural extracts for tumor cells, and the IHC staining confirmed that these extracts regulate the expression of proteins related to cell proliferation, apoptosis, and metastasis. In summary, natural extracts have the potential to inhibit melanoma cell proliferation, induce apoptosis, and prevent metastasis, indicating great potential for melanoma treatment.