研究不同植物屬別之粗萃物對大腸直腸癌的抑癌作用

大腸直腸癌是全世界最常見也是最主要的癌症死亡原因之一，當患者被診斷罹患大腸直腸癌時。一般治療通常包括手術切除並輔以化學療法。然而，在臨床上常見的化療藥物，由於腫瘤細胞與正常細胞之間的選擇性低而導致許多副作用。因此，開發出對於人體危害較小，且可用於替代目前治療大腸直腸癌的新型化療藥物還是有其需求性。在先前文獻中，已經證實沒藥 (Commiphora Myrrha, CMy) 可以誘導肺癌，胰腺癌，乳腺癌和前列腺癌細胞凋亡，然而對大腸直腸癌的抗腫瘤作用仍然未知。另外，已證明鋪地百里香 (Thymus serpyilum, TSe) 可以誘導乳癌細胞凋亡。因此，本篇研究旨在探討沒藥和鋪地百里香粗萃物在體外和體內實驗中對大腸直腸癌的抗癌作用及其機制。首先以MTT assay及流式細胞儀的分析，發現CMy和TSe粗萃物可抑制大腸直腸癌細胞的生長，並使HT-29細胞停滯在G0/G1期，同時會使Sub-G1期上升。另外，根據TUNEL測定的結果，經過CMy和TSe粗萃物作用，可在HT-29細胞中觀察到細胞凋亡形態。接著透過西方點墨法及Caspase活性測試，證實CMy和TSe粗萃物會活化外在及內在細胞凋亡路徑。此外，CMy和TSe粗萃物在wound healing assay、transwell assay及 Colony formation assay中均能抑制大腸癌腸癌細胞的移行作用及轉移後的生長能力。最後在動物治療模式中，顯示CMy和TSe粗萃物可抑制大腸直腸癌腫瘤的生長，並且對於正常器官幾乎沒有毒性。除此之外，通過免疫組織化學染色，在CMy和TSe粗萃物治療的組別中，活化態Caspase-3和PARP的表達量增加，而PCNA和MMP9的表達減少。總合上述實驗，證明CMy和TSe粗萃物對於大腸直腸癌細胞具有抑癌的效果，並有可能作為一種新型的大腸直腸癌化療藥物。

Colorectal cancer (CRC) is one of the most common and the leading cause of cancer mortality worldwide. The standard treatment for patients who are diagnosed with CRC usually involves surgical removal in combination treatment with chemotherapy. However, the classic chemotherapy drugs lead to adverse side effects because of low selectivity between tumor and normal cells. Therefore, it is important to develop novel drugs for alternative chemotherapeutic agents with less harmful to human body in CRC therapy. In previous studies, Commiphora myrrha (CMy) has shown that CMy could induce cell apoptosis in lung, pancreatic, breast and prostate cancer, while the anti-tumor effect for CRC is still unknown. In addition, Thymus serpyilum (TSe) has been proved that TSe induced apoptosis in breast cancer. Therefore, this study aimed to explore the anti-tumor effects and mechanisms of CMy and TSes on CRC cell line in vitro and in vivo. In in vitro studies, we found that CMy and TSes reduced proliferation of CRC cells, arresting HT-29 cells at G0/G1 phase and presented elevation in Sub-G1 phase. Additionally, according to the result of TUNEL assay, HT-29 cells showed cell apoptotic morphology after treatment with CMy and TSes. Western blot analysis and caspase activity assay revealed that the extrinsic and intrinsic apoptosis related proteins were activated by both extracts. In addition, CMy and TSes inhibited the migration of CRC cells in wound healing assay and transwell assay and proliferation after migration in Colony formation. In in vivo study, CMy and TSes suppressed the growth of CRC tumor and had lower or no toxicity in tumor-bearing mice. In addition, the expression of cleaved Caspase-3 and PARP were increased, while PCNA and MMP9 were decreased after CMy and TSes treatment via detection of IHC staining. Taken together, our study proved that CMy and TSes exerted anti-tumor activity in CRC cells and potential in development as a novel anti-tumor agent for CRC therapy.