Whole genome gene expression changes and hematological effects of rikkunshito in patients with advanced non-small cell lung cancer receiving first line chemotherapy

Abstract

It has been demonstrated that the traditional Chinese medicine rikkunshito, ameliorates anorexia in several types of human cancer and attenuates lung injury by inhibiting neutrophil infiltration. The current study investigated the clinical and hematological effects of rikkunshito and its underlying mechanisms of action in the treatment of advanced non-small cell lung cancer (NSCLC). The Illumina microarray BeadChip was used to analyze the whole-genome expression profiles of peripheral blood mononuclear cells in 17 patients with advanced NSCLC. These patients were randomized to receive combination chemotherapy (cisplatin and gemcitabine) with (n=9, CTH+R group) or without (n=8, CTH group) rikkunshito. The primary endpoint was the treatment response and the categories of the scales of anorexia, nausea, vomiting and fatigue; secondary endpoints included the hematological effect and whole genome gene expression changes. The results of the current study indicated that there were no significant differences in clinical outcomes, including treatment response and toxicity events, between the two groups. Median one-year overall survival (OS) was 12 months in the CTH group and 11 months in the CTH+R group (P=0.058 by log-rank test), while old age (>60 years old) was the only independent factor associated with one-year OS (hazard ratio 1.095, 95% confidence interval, 1.09-1.189, P=0.030). Patients in the CTH+R group experienced significantly greater maximum decreases in both white cell count (P=0.034) and absolute neutrophil count (P=0.030) from the baseline. A total of 111 genes associated with neutrophil apoptosis, the cell-killing ability of neutrophils, natural killer cell activation and B cell proliferation were up-regulated following rikkunshito treatment. A total of 48 genes associated with neutrophil migration, coagulation, thrombosis and type I interferon signaling were down-regulated following rikkunshito treatment. Rikkunshito may therefore affect the blood neutrophil count when used with combination chemotherapy in patients with NSCLC, potentially by down-regulating prostaglandin-endoperoxidase synthase 1, MPL, AMICA1 and junctional adhesion molecule 3, while up-regulating elastase, neutrophil expressed, proteinase 3, cathepsin G and cluster of differentiation 24.

Keywords: chemotherapy; microarray gene expression; neutropenia; non-small cell lung cancer; rikkunshito.

Figure 1.

One-year survival analysis for the patients with lung cancer treated with CTH alone (CDDP+GEM) or CTH + rikkunshito. (A) Kaplan-Meier estimates of 1-year progression free survival (P=0.390, Log-Rank test). (B) Kaplan-Meier estimates of 1-year overall survival (P=0.058, Log-Rank test). (C) Kaplan-Meier estimates demonstrated that patients ≤60 years old had a better 1-year survival rate than those >60 years old (P=0.015, Log-Rank test). CTH, cisplatin (CDDP)-based chemotherapy; CDDP, cisplatin; GEM, gemcitabine.

Figure 2.

Three-year survival analysis for the patients with lung cancer patients treated with CTH alone (CDDP+GEM) or CTH + rikkunshito. (A) Kaplan-Meier estimates of 3-year progression free survival (P=0.591, Log-Rank test). (B) Kaplan-Meier estimates of 3-year overall survival (P=0.261, Log-Rank test). CTH, cisplatin (CDDP)-based chemotherapy; CDDP, cisplatin; GEM, gemcitabine.

Figure 3.

Rikkunshito-regulated signaling pathways. Microarray-identified signaling pathways enriched following the addition of rikkunshito in the chemotherapy regimen for the advanced non-small cell lung cancer patients. The majority of these genes targeted insulin, cyclin-dependent kinase 1, Grb-2-related adaptor protein 2 and interleukin-2.