The International Agency for Research on Cancer reported that in 2020, approximately 10.1 million deaths due to cancer occurred worldwide, with liver cancer identified as the third leading cause of death, resulting in around 830,000 deaths [1]. Hepatocellular carcinoma (HCC) was identified as the most common form of primary liver cancer, making it the most prevalent type [2]. The specificity of cancer necessitates the adoption of different strategies for the diagnosis and treatment of each type. It is also noted that some cancers evade immune surveillance, resulting in stronger immune responses that are associated with treatment failures [3,4]. Evaluating the mechanisms of cancer cell movement and growth allows for the assessment of their motility and invasion processes, providing important insights for the development of new therapeutic strategies [[5], [6], [7]].

The water extract of Phellinus Linteus (PLWE), a common traditional Chinese medicine, is rich in various active components, such as polysaccharides and phenolic compounds, and has a long history of medicinal use [8]. Phellinus Linteus has been found to possess antioxidant properties that reduce oxidative stress and improve cardiovascular health. Studies have indicated that it not only improves insulin sensitivity and lowers blood glucose levels but also alleviates complications through its anti-inflammatory effects [9]. Furthermore, Phellinus Linteus is noted to enhance immune function and promote antitumor immune responses [10]. Specifically, its primary mechanisms in inhibiting tumor cell proliferation and inducing apoptosis include regulating the cell cycle and suppressing the migration and invasion capabilities of tumor cells [11].

The mechanical changes in cells are closely associated with the occurrence and development of cancer [12,13]. Cancer mechanisms have been primarily investigated through the observation of tumor slices under a microscope or by establishing in vitro models for drug screening [14]. The main aims of anticancer drugs are to inhibit the growth and spread of cancer cells while minimizing damage to normal cells [15,16]. Studies have shown that the motility of cancer cells is evident not only in cell migration and intercellular interactions but also in their remarkable deformability within confined spaces [17,18]. Calzado-Martin et al. simultaneously detected three breast cell lines using multiparametric Atomic Force Microscopy (AFM), obtaining morphological and stiffness images that demonstrate the significant contribution of actin to cell stiffness [19]. Additionally, Rigato et al. employed oscillatory AFM probes to measure the viscoelastic properties of cells over short periods, revealing that the variation patterns of normal and malignant cancer cells differ at high frequencies [20]. This research suggests a potential link between the softness of cancer cells and a decrease in the surface tension of their membranes, which may enhance their deformability. The close relationship between mechanical properties and biological behavior emphasizes the necessity of examining the response of cancer cells to various stress conditions to evaluate their performance in the invasion process [21]. Omidvar et al. compared the adhesion of three breast cancer cell lines (MCF-7, T47D, and MDA-MB-231) with differing metastatic potentials, revealing a negative correlation between cell adhesion and their invasive capabilities [22]. Measuring the stiffness and other mechanical properties can uncover the distinctions between cancer and normal cells, facilitating a better understanding of the behavior and mechanisms of cancer [[23], [24]]. While cancer treatment is becoming more individualized, the rapid proliferation and migration of cancer cells still pose significant challenges in tumor formation. The mechanical characteristics of cancer and their complex pathological mechanisms significantly contribute to treatment failures [25,26].

The survival and proliferation of cancer cells, along with their complex pathogenesis, are the main reasons for treatment failure [27]. AFM has been utilized to achieve real-time imaging of cell surface morphology and to obtain mechanical characteristics, which assist in evaluating cellular status and treatment effectiveness [[28], [29]]. In this study, a water extraction method was employed to obtain the extract from Phellinus Linteus and assess its impact on the state of SMCC-7721 cells over various treatment durations. The SMCC-7721 cells were treated with Phellinus Linteus water extract (PLWE), followed by cell viability assays. Scanning electron microscopy (SEM) and AFM were utilized to assess the multidimensional parameters of the cells over various treatment durations, illustrating their dynamic changes. The significance of this research lies in investigating how SMCC-7721 cells respond to PLWE, assessing its potential therapeutic effects in cancer treatment, and supporting its application in tumor therapy.