Noscapine and Apoptosis in Breast and Other Cancers

Abstract

Breast cancer is the second leading contributor to the age-standardized mortality rate, for both sexes and all ages worldwide. In Europe and the United States, it is the second leading cause of mortality, with an incidence rate of about 2.6 million cases per year. Noscapine, a well-known alkaloid used as a cough suppressant, demonstrated anti-tumor effects by triggering apoptosis in various cancer cell lines and has the potential to become another ally against breast, ovarian, colon, and gastric cancer, among other types of malignancy. Apoptosis plays a crucial role in the treatment of cancer. Noscapine affected BAX, CASP8, CASP9, NFKBIA, and RELA gene and protein expression in the MCF-7 and MDA-MB-231 cell lines. Gene expression was higher in tumor than in normal tissue, including the BAX expression levels in lung, ovary, endometrium, colon, stomach, and glioblastoma patients; BCL2L1 expression in endometrium, colon, and stomach patients; CASP8 gene expression levels in lung, endometrium, colon, stomach, and glioblastoma patients; RELA in colon, stomach, and glioblastoma patients; and NFKBIA in glioblastoma patients. It can be concluded that noscapine affected genes and proteins related to apoptosis in cancer cell lines and several types of cancer patients.

Keywords: Bax; Bcl-2; apoptosis; breast cancer; caspase-8; caspase-9; noscapine.

Figure 1

Structures of (A) noscapine, (B) cyclic ether fluorinated noscapine analog (CEFNA), (C) 9-Cl-noscapine (EM015), (D) 9-Br-noscapine, and (E) 9-I-noscapine.

Figure 2

(A) The heatmap table shows the correlations between TP53, the tumor protein p53 gene, and BAX, the BCL2-associated X gene; BCL2L1, the BCL2-like 1 gene; CASP8, the caspase-8 gene; CASP9, the caspase-9 gene; NFKBIA, the NFKB inhibitor alpha gene; and RELA, the RELA proto-oncogene (NFkB, Rel A) in breast invasive carcinoma (BRCA) subtypes. The red color indicates a statistically significant positive correlation (Spearman’s, p < 0.05) and gray denotes a non-significant result. (B) Box plots show significant correlations between TP53 expression with purity adjustment (left) and CASP8, CASP9, and RELA gene expression levels (right) in BRCA subtypes. Correlation values for each analysis are stated in red on the right (Spearman’s, p < 0.05). Raw data were extracted from TIMER2.0 (http://timer.cistrome.org), accessed on 14 September 2023 [59].

Figure 3

Differential gene expression levels between tumor and normal tissue in breast invasive carcinoma (BRCA). The box plots show the distribution of gene expression levels of (A) BAX, the BCL2-associated X gene; (B) BCL2L1, the BCL2-like 1 gene; (C) CASP8, the caspase-8 gene; (D) CASP9, the caspase-9 gene; (E) RELA, the RELA proto-oncogene; and (F) NFKBIA, the NFKB inhibitor alpha gene in tumor versus normal tissue (Wilcoxon rank-sum test, ***: p < 0.001). Raw data were extracted from the Tumor Immune Estimation Resource database v 2.0 (TIMER2.0, http://timer.cistrome.org), accessed on 14 September 2023 [59]. (1) BRCA. Tumor (n = 1093). (2) BRCA. Normal (n = 112). (3) BRCA-Basal. Tumor (n = 190). (4) BRCA-Her2. Tumor (n = 82). (5) BRCA-LumA. Tumor (n = 564). (6) BRCA-LumB. Tumor (n = 217).

Figure 4

Xena Chart View showing box plot transcript expression of (A) BAX, the BCL2-associated X gene; (B) BCL2L1, the BCL2-like 1 gene; (C) CASP8, the caspase-8 gene; (D) CASP9, the caspase-9 gene; (E) RELA, the RELA proto-oncogene; and (F) NFKBIA, the NFKB inhibitor alpha gene in breast invasive carcinoma (BRCA). Cohort: TCGA Breast Cancer (BRCA), n = 782, stratified by nature2012 for estrogen receptor status (ER) (one-way ANOVA, p < 0.05). Raw data were extracted from the University of California, Santa Cruz (ena.ucsc.edu) UCSC Xena functional genomics explorer, accessed on 14 September 2023 [60].

Figure 5

(A) Correlations between TP53, the tumor protein p53 gene, and BAX, the BCL2-associated X gene; BCL2L1, the BCL2-like 1 gene; CASP8, the caspase-8 gene; CASP9, the caspase-9 gene; NFKBIA, the NFKB inhibitor alpha gene; and RELA, the RELA proto-oncogene (NFkB, Rel A) in different types of cancer. The red color indicates a statistically significant positive correlation (Spearman’s, p < 0.05), blue indicates a statistically significant negative correlation (Spearman’s, p < 0.05), and gray denotes a non-significant result. (B). Box plots show significant correlations between TP53 expression with purity adjustment (left) and BAX, BCL2L1, CASP8, CASP9, RELA, and NFKBIA gene expression levels (right) in different types of cancer. Correlation values for each analysis are stated in red on the right (Spearman’s, p < 0.05). Raw data were extracted from TIMER2.0 (http://timer.cistrome.org), accessed on 10 January 2024 [59]. TCGA abbreviations—LUAD: lung adenocarcinoma; LUSC: lung squamous cell carcinoma; OV: ovarian serous; UCEC: uterine corpus endometrial carcinoma; COAD: colon adenocarcinoma; STAD: stomach adenocarcinoma; GBM: glioblastoma multiforme.

Figure 6

Differential gene expression levels between tumor and normal tissue in different types of cancer. The box plots show the distribution of the gene expression levels of (A) BAX, the BCL2-associated X gene; (B) BCL2L1, the BCL2-like 1 gene; (C) CASP8, the caspase-8 gene; (D) CASP9, the caspase-9 gene; (E) RELA, the RELA proto-oncogene; and (F) NFKBIA, the NFKB inhibitor alpha gene in tumor versus normal tissue (Wilcoxon rank-sum test, *: p < 0.05, **: p < 0.01, ***: p < 0.001). Raw data were extracted from the Tumor Immune Estimation Resource database v 2.0 (TIMER2.0, http://timer.cistrome.org), accessed on 14 September 2023 [59]. (a) LUAD. Tumor (n = 515). (b) LUAD. Normal (n = 59). (c) LUSC. Tumor (n = 501). (d) LUSC. Normal (n = 51). (e) OV. Tumor (n = 303). (f) UCEC. Tumor (n = 545). (g) UCEC. Normal (n = 35). (h) COAD. Tumor (n = 457). (i) COAD. Normal (n = 41). (j) STAD. Tumor (n = 415). (k) STAD. Normal (n = 35). (l) GBM. Tumor (n = 153). (m) GBM. Normal (n = 5).

Scheme 1

(A) Noscapine causes changes in apoptotic genes in cancer in human patients, such as glioblastoma, lung, endometrium, breast, stomach, ovary, and colon. The red arrow indicates an increase and the blue arrow indicates a decrease in gene expression. (B) It also causes changes in apoptotic genes and protein expression in breast cancer cell lines.