As one of the most common cancers in the world, colorectal cancer (CRC) is also a major contributor to cancer-related death. Patients with colorectal cancer still have a poor prognosis, with a high risk of metastasis and recurrence, despite major improvements in surgical procedures, chemotherapy, and targeted medicines. Thus, identifying novel therapeutic agents and strategies becomes crucial to enhance the clinical outcomes for CRC patients [[1], [2], [3]].

Alpinia officinarum Hance (A. officinarum), widely known as galangal, is a perennial herb that falls under the Zingiberaceae family. This plant is commonly found in Southeast Asia and is grown in Vietnam and southern China, and its rhizome has been used as a spice in Europe for over 1000 years. It grows in the wild and is also grown as food in Guangdong Province, China [4]. It is reported that galangal extract exhibits various biological activities, including antioxidant, anti-inflammatory, antibacterial, antidiabetic, neuroprotective, and liver-protective effects [5]. Recently, many studies suggested that galangal extract has anti-cancer biological activity, such as liver cancer, breast cancer, gastric cancer, etc. However, the potential bioactive compound of galangal as well as the molecular mechanism of its anti-colorectal effect is still need to be further explored.

Apoptosis, cell cycle arrest, and production of reactive oxygen species (ROS) are the main mechanisms underlying the anticancer effects of many natural compounds. Apoptosis, a type of programmed cell death, is governed by the intricate balance between pro-apoptotic and anti-apoptotic factors. Cell cycle progression is meticulously regulated by a series of checkpoints, and any disruption can result in unchecked cell proliferation. ROS serve as signaling molecules in small amounts, and excessive accumulation can lead to oxidative stress, causing DNA damage and cell death. The fate of cancer cells and cellular homeostasis rely on maintaining a balance among these processes [[6], [7], [8]].

The MAPK pathway plays a significant role in the tumorigenesis and disease progression of CRC. When activated, this pathway can lead to increased cell proliferation, survival, and metastasis, whereas its inhibition can induce apoptosis and cell cycle arrest [9]. For instance, recent studies indicate that inhibiting the MEK/ERK pathway can substantially decrease CRC cell viability and induce apoptosis by regulating pro-apoptotic proteins such as Bcl-2 and Bax. Moreover, activation of this pathway is associated with ROS level regulation, which can affect cell fate decisions. High ROS levels can cause oxidative stress and DNA damage, resulting in apoptosis, whereas the MEK/ERK pathway can control ROS production to preserve cellular homeostasis. Nonetheless, the role of the MAPK pathway in galangal's anti-cancer effects in CRC remains unknown [10,11].

In this study, we identify the potential bioactive compounds of galangal using network pharmacology, explore the related mechanism of bioactive compound of galangal anti-colorectal cancer by conducting RNA sequencing analyses, then carry out a series of studies to verify these mechanisms. The whole study flowchart can be seen in Fig. 1.