Cancer Letters

Cancer Letters

Volume 268, Issue 1, 8 September 2008, Pages 166-175
Cancer Letters

Anticancer effect of tetrandrine on primary cancer cells isolated from ascites and pleural fluids

https://doi.org/10.1016/j.canlet.2008.03.059Get rights and content

Abstract

Objective

The present study was designed to examine the effect of tetrandrine on primary cancer cells isolated from the ascites or pleural fluids of patients with metastatic cancers.

Methods

Primary cancer cells were isolated from the pleural fluids (n = 13) or ascites (n = 21) of patients. Compared with culture cell lines, the response of these cancer cells to tetrandrine and chemotherapeutic agents commonly used in clinical practice were determined by WST-8 assay. Tetrandrine-induced apoptosis in primary cancer cells was determined by Annexin V-FITC assay. Quantitative RT-PCR was used to examine the role of apoptotic associated genes in the anticancer effect of tetrandrine.

Results

The primary cancer cells isolated from effusions showed sensitivity to tetrandrine with IC50 values of 38.23 ± 25.77 μM, similar to the IC50 in established cell lines. Patients with gastric cancers were more sensitive to tetrandrine than patients with lung cancers (P = 0.04). Four cancer cells isolated from effusions were resistant to tetrandrine, which also had increased tolerance to docetaxel, cisplatin and 5-fluorouracil. We also observed a weak but significant correlation between sensitivity to tetrandrine and cellular expression of bcl-2 (P = 0.035, r = −0.364).

Conclusions

Using cancer cells isolated from the ascites or pleural fluids, this study shows the potential anticancer effect of tetrandrine against primary cancer cells.

Introduction

Malignant effusion is a common and distressing complication of advanced malignant diseases. Approximately half of patients with metastatic cancers develop malignant pleural effusion or ascites at some point, which is likely to cause significant symptoms such as dyspnea and abdominal distention [1]. Tumor spread via survival and proliferation of tumor cells in body cavity fluid is an important route of metastasis and a frequent cause of morbidity in many malignancies. Evacuation of the pleural fluid and prevention of its re-accumulation are the main goals of management. However, optimal treatment is controversial and there is no universally standard approach. Development of novel methods and potent drugs to control malignant pleural effusion should be a high priority in palliative care of cancer patients.

Tetrandrine (TET), a bis-benzylisoquinoline alkaloid isolated from the Chinese herb “Han-Fang-Chi” (Stephania tetrandra S. Moore), has been reported to have many pharmacological effects including anti-inflammation [2], antioxidant [3], antifibrotic [4] and anti-tumor activity [5]. Tetrandrine regulates cellular redox states, which may reduce proliferation and induce apoptosis in tumor cells [6]. It also induces cell cycle arrest in the G1 phase via p53 and the Fas/FasL apoptotic system in HepG2 cells [7]. Furthermore, it has been shown to be a potent inhibitor of P-gp drug efflux and a resistant reversal agent to daunorubicin, vinblastine and doxorubicin in leukemia cells [8]. Our earlier study has demonstrated synergistic interaction between tetrandrine and chemotherapeutic agents in human gastric cancer cell lines [9].

To date, studies demonstrating the anti-tumor activity of tetrandrine have mainly been done with established cell lines. In this study, we report the cytotoxicity effect of tetrandrine on primary cancer cells isolated from ascites and pleural fluid of patients. To elucidate the mechanisms possibly involved, the expression levels of apoptosis associated genes were also determined.

Section snippets

Cell lines and cell culture

Human gastric cancer cells BGC-823, liver cancer cells SMMC-7721, lung cancer cells A549 and colon cancer cells Lovo were obtained from Shanghai Institute of Cell Biology (Shanghai, China). These cells were grown in RPMI 1640 medium (GIBCO BRL) with 10% fetal bovine serum (FBS) and 100 U/mL penicillin–streptomycin at 37 °C in a water-saturated atmosphere with 5% CO2.

Chemotherapeutic agents

The chemotherapeutic drugs (docetaxel, cisplatin, 5-fluorouracil) used in the assay were obtained as vials for injection and made up

Sensitivity of cancer cells to tetrandrine

We first examined the cytotoxicity of each drug for BGC-823, SMMC-7721, A549 and Lovo cells lines. Table 1 shows the IC50 doses for these four cells following exposure to tetrandrine or chemotherapeutic agents. Consistent with the studies that have been done on established cell lines, we found that a 3-day treatment with 10–40 μM tetrandrine inhibited ≈50% of the cells in several cell lines. The IC50 values of tetrandrine for BGC-823, SMMC-7721, A549 and Lovo cells were 11.20 μM, 21.76 μM, 24.0 μM

Discussion

Effusions may be the presenting sign of malignancy or the first manifestation of recurrence and relapse. Here, we first describe the use of cancer cells isolated from effusions for studying responses to tetrandrine and other anticancer agents used currently in clinical. These primary cancer cells would be a more accurate model for initial drug trails, as they reflect the situation seen in the body. In our studies of the response of primary cancer cells to tetrandrine, we made several

Acknowledgments

This work is supported in part by Health Department in Jiangsu Province (H200640), National Nature Science Foundation of China (30670958) and Medical Technology Development Foundation of Nanjing (ZKX05015).

References (25)

  • Y.J. Chen

    Potential role of tetrandrine in cancer therapy

    Acta Pharmacol. Sin.

    (2002)
  • Y. Dong et al.

    In vitro inhibition of proliferation of HL-60 cells by tetrandrine and coriolus versicolor peptide derived from Chinese medicinal herbs

    Life Sci.

    (1997)
  • Cited by (39)

    • Tetrandrine induces apoptosis in human neuroblastoma through regulating the Hippo/YAP signaling pathway

      2019, Biochemical and Biophysical Research Communications
      Citation Excerpt :

      TET has been proved to hold potential therapeutic value as it bears a variety of anti-inflammatory, anti-hypertensive, anti-arrhythmic, and anti-allergenic properties [6]. Recent studies have confirmed the anti-cancer activity of TET in several different cancers [7,8]. TET was shown to induce cell death and autophagy in human oral cancer cells and gastric cancer cells [9,10].

    • Tetrandrine and cancer – An overview on the molecular approach

      2018, Biomedicine and Pharmacotherapy
      Citation Excerpt :

      Tetrandrine mediated caspase activation and cleavage of PARP by caspase -3 proteins to bring about DNA fragmentation was observed in hepatic stellate cells [70]. As it has already been observed for many anticancer drugs [71,72], tetrandrine brings about enhanced activity of intrinsic caspase pathway with an elevated intracellular ROS concentration under in vitro and in vivo conditions [73–78]. Perhaps, the ability of tetrandrine to upregulate BAX and BID might involve the release of Cyt C from mitochondria to induce apoptosis.

    • Plant-Derived Prooxidants as Potential Anticancer Therapeutics

      2016, Studies in Natural Products Chemistry
    • Spectroscopic studies on the interaction between tetrandrine and two serum albumins by chemometrics methods

      2013, Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy
    • Taxol-oligoarginine conjugates overcome drug resistance in-vitro in human ovarian carcinoma

      2012, Gynecologic Oncology
      Citation Excerpt :

      By testing the cancer cells without selecting for growth advantages or expanding particular subpopulations, the assay emulates the heterogeneity of the clinical disease. The growth rate of primary tumor samples (ascites cells and dissociated solid tumor cells) in vitro replicates the physiological environment where cancer cells grow more slowly than established cell lines [19,20]. The diverse set of transporter-drug conjugates evaluated includes prodrugs that exhibit extended stabilities under assay conditions, allowing them to be administered and remain intact until cell entry, after which they are rapidly cleaved in the intracellular reducing environment, releasing the free drug at tunable rates ranging from minutes to hours and controlled by linker design.

    View all citing articles on Scopus
    1

    These authors contributed equally to this work.

    View full text