OncoWuXi Express: Paclitaxel-Induced Resistant Tumor Models
Introduction：
OncoWuXi Express will continue to keep you informed about updates to our online tumor model database (OncoWuXi Database), as well as our recent progress in cancer and autoimmune research. In this issue, we would like to share with you the newly established paclitaxel resistant HGC27 and HCC1806 tumor models.
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Paclitaxel, known for its high effectiveness and low toxicity, is a widely utilized natural anticancer drug with broad-spectrum applications. It is commonly employed in clinical treatments of various malignant tumors including breast cancer, ovarian cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, and gastric cancer, making it one of the most common chemotherapy drugs. Paclitaxel stabilizes microtubules by amplifying the binding of microtubule protein dimers and organizing their depolymerization, thereby inhibiting the normal dynamic reorganization of microtubules, which is pivotal for the functionality of cells during interphase and mitosis. This process leads to cell cycle arrest at the G2/M phase, triggering irregular or halted mitosis, impeding tumor cell replication, and barring the division of cancer cells, thereby ultimately inducing apoptosis. Paclitaxel also has the ability to trigger a variety of signaling pathways, with its action mechanism seemingly related to the initiation of pro-apoptotic signals (Figure 1) [1,2]. However, with prolonged clinical use, the inevitable occurrence of paclitaxel resistance hampers the progress of cancer treatment. Paclitaxel resistance involves multiple mechanisms, among which the biological pathway targets and genes associated with resistance include ABCB1, TUBB, BCL-2, and more. (Figure 2) [3]. Therefore, gaining a better understanding of the molecular mechanisms that underlying the reversal of resistance, in conjunction with enhancing drug efficacy and utilizing targeted inhibition or specific drug delivery methods, is of paramount importance for optimizing resistance prevention or reversal and enhancing the efficacy of chemotherapy.
Figure 1. Action mechanism of paclitaxel (PTX) [1]
Figure 2. Paclitaxel resistance-related genes [2]
To facilitate the discovery and development of new drugs, WuXi Biology has established gastric cancer HGC27 (Paclitaxel-R-HGC27) and breast cancer HCC1806 (Paclitaxel-R-HCC1806) cell lines that are stably resistant to paclitaxel, through long-term drug administration in vitro and in vivo. We have also established corresponding in vivo tumor models for these two resistant cell lines.
The Paclitaxel-R-HGC27 cell line was derived from parental HGC27 cells via long-term in vitro drug treatment. In vitro CTG (CellTiter-Glo) assay demonstrated that Paclitaxel-R-HGC27 possesses a pronounced paclitaxel-resistant phenotype compared to parental HGC27 (Figure 3A and B). Subsequently, Paclitaxel-R-HGC27 cells were subcutaneously xenografted into immunodeficient mice, exhibiting a certain degree of resistance to both 15 mg/kg and 30 mg/kg dosages of paclitaxel, compared to parental HGC27 cells (Figure 3C). Recent studies indicate that members of the ATP Binding Cassette (ABC) transporter protein family can facilitate drug efflux, prompting an increased expulsion of various anticancer drugs from tumor cells. Consequently, the upregulation of expression and activity of ABC transporter proteins represent a classical mechanism of chemotherapeutic resistance[4]. Flow cytometry analysis of Paclitaxel-R-HGC27 cells showed upregulation in both ABCB1 (ATP Binding Cassette subfamily B member 1) and ABCG2 (ATP Binding Cassette super-family G member 2) proteins (Figure 3D).
Figure 3. The establishment of paclitaxel resistant HGC27 cell line (A) The effect of paclitaxel in the parental and resistant cell lines examined by the in vitro CTG assay. (B) IC50 values of parental and resistant cell lines treated with paclitaxel. (C) Tumor growth curves of parental and resistant cell line xenograft models treated with paclitaxel in vivo. (D) ABCB1 and ABCG2 expression in parental and resistant cell lines detected by flow cytometry.
Unlike the establishment strategy of Paclitaxel-R-HGC27, Paclitaxel-R-HCC1806 was obtained through long-term drug induction in HCC1806-bearing mice. When a resistant phenotype was observed, the tumor tissue was harvested and subsequently processed into single cells in vitro. The CTG results showed that Paclitaxel-R-HCC1806 exhibited paclitaxel resistance compared to parental HCC1806 (Figure 4A and B). Following this, Paclitaxel-R-HCC1806 cells were subcutaneously xenografted into immunodeficient mice and treated with paclitaxel at a dosage of 25 mg/kg. Compared to parental cells, the resistant cells exhibited significant resistance (Figure 4C).
Figure 4. The establishment of paclitaxel- resistant HCC1806 cell line (A) The effect of paclitaxel in the parental and resistant cell lines examined by the in vitro CTG assay. (B) IC50 values of parental and resistant cell lines treated with paclitaxel. (C) Tumor growth curves of parental and resistant cell lines treated with paclitaxel in vivo.
Conclusion
These results have demonstrated that we have successfully established two paclitaxel-induced, drug-resistant tumor cell lines and in vivo models. Both models can be ideal tools for exploring new therapies aimed at overcoming drug resistance.
References:
[1] Kampan, Nirmala Chandralega et al. “Paclitaxel and Its Evolving Role in the Management of Ovarian Cancer.” BioMed research international vol. 2015 (2015): 413076. doi:10.1155/2015/413076
[2] Maloney, Sara M et al. “Mechanisms of Taxane Resistance.” Cancers vol. 12,11 3323. 10 Nov. 2020, doi:10.3390/cancers12113323
[3] Dorman, Stephanie N et al. “Genomic signatures for paclitaxel and gemcitabine resistance in breast cancer derived by machine learning.” Molecular oncology vol. 10,1 (2016): 85-100. doi:10.1016/j.molonc.2015.07.006
[4] Chew SC, Singh O, Chen X, et al. The effects of CYP3A4, CYP3A5, ABCB1, ABCC2, ABCG2 and SLCO1B3 single nucleotide polymorphisms on the pharmacokinetics and pharmacodynamics of docetaxel in nasopharyngeal carcinoma patients. Cancer Chemother Pharmacol. 2011;67(6):1471-1478. doi:10.1007/s00280-011-1625-9
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