A Review of The Role of Hemoglobin in Different types of Cancer ISSN 2753-8176 (online), DOI:10.13140/RG.2.2.24610.20164

Ana Pedro1

1Gwyntwr1386 Pharmacy, Regus Chester Business Park, Heronsway, Chester, CH49QR, UK

Keywords:

1. Corresponding author: info@gwyntwr1386.com

A role for haemoglobin in cancer diagnosis in general is supported by a good number of publications. A previous work indicated the possibility of a direct relationship between the hemoglobin level and not only the early stages of cancer, but also the chemically induced precancerous condition (1). Interestingly, in patients with a diagnosis of primary lung, colorectal, breast or liver cancer, higher post-diagnosis haemoglobin change regardless of the baseline haemoglobin levels and the direction of changes associates with a significantly shorter survival (2).

Here we summarize, the results of our findings in what concerns proteomics analyzis of several different cancer patient samples and of cancer cell lines and cancer models for several different hemoglobin subunits and variants.

Breast cancer

Preliminary data of our lab have showed that we have identified a biomarker for early stage ER+ breast cancer, HCG1745306 isoform CRA-a (3).Additional published data from our lab shows that hemoglobin subunit alpha (HBA1), which was found to be present in urine samples of patients with early breast cancer (BC) (4), also presents very high mascot scores in plasma samples of patients with early BC. The mascot scores, however, drop in the plasma of patients with locally advanced BC and of patients with certain metastatic patterns. Also, HBA1 is not significantly present in the primary BC tumor tissues, although bone metastases, show a high score for this protein. Hemoglobin subunit beta (HBB), which was found to increase breast cancer cells aggressiveness and associates with poor prognosis (5), also presents very high mascot scores in plasma samples of patients with early BC. The mascot scores, however, drop in the plasma of patients with locally advanced BC, but are significant in the primary BC tissues what allows us to think might be secreted by the tumor. Bone metastases, also show a high score for this protein (6).

Melanoma

We found that the mascot scores, for hemoglobin subunits alpha (HBA), delta (HBD) and gamma (HBG) increase considerably for the melanoma patients, both non-metastatic and metastatic, in comparison to healthy patients (controls). Also, melanoma patients with undetected metastasis or that will develop metastasis in the future might express HBB and a fragment of hemoglobin epsilon (HBE1) (7).

Pancreatic cancer

We found out that normal mouse pancreas tissue conditioned media (CM) does not show any hemoglobin subtype. However, PanIn CM may show good amount of HBB and vestigial amounts of HBA. By other side, AsPc1 CM shows good amounts of both types of hemoglobin subunits. The remaining samples only show hemoglobin subunit alpha: BXPC3 CM shows vestigial amounts, HPAF2 CM vestigial amounts or a good amount, MiaPaca2 CM a good amount, PAN02 CM a good amount or nothing and PANC1 CM nothing. These results suggest that in pancreatic cancer, similarly to what might happen with other cancer types (5), HBB might play a role, in particular in the initial stages of pancreatic cancer. HBA might have a role in established PC.

Colorectal cancer

We found out that pre-malignant tissues such as human colon epithelial cells express different hemoglobin subunits. In particular, HBB which possibly is involved in tumor progression (5) compared with pre-malignant colon tissue presents a raised score in primary tumor cells lines. Curiously, when P53 is overexpressed the score goes down again. The score is bigger again at stage III cell line, SW620. We can see moreover, HBB scores raises when comparing the primary tumor with the p53 knock-out cell line. By adding vitamin D or by adding vitamin D and P53 overexpression the scores for HBB go down again. We found out that HBE is only expressed by the primary tumor and by the P53 knock-out cell line where there is an huge increase in HBE score compared with the primary tumor. HBG is expressed in the pre-malignant colon tissue and in the primary cancer cell lines which show an increased score in comparison to the pre-malignant tissues. Moreover, the scores for this hemoglobin are slightly raised in p53 knockout and decrease with adding vitamin D. The human alpha globin gene cluster is located on chromosome 16. The alpha-2 and alpha-1 coding sequences are identical. Two alpha chains plus two beta chains constitute HbA, which in normal adult life comprises about 97% of the total hemoglobin; alpha chains combine with delta chains to constitute HbA-2, which with fetal hemoglobin makes up the remaining 3% of adult hemoglobin. HBA is expressed by pre-malignant tissues and its score raises in the primary cancer tissues. P53 overexpression and in stage III tissues there are slight decreases in this protein score. This protein scores raises in the p53 knockout in comparison to the primary cancer, however adding vitamin D or vitaminD+p53 overexpression do slightly increase the scores for this protein. In conclusion, all hemoglobin subunits (gamma 2, beta and alpha) are expressed in pre-malignant colon tissues except hemoglobin subunit epsilon. All hemoglobins scores are raised and hemoglobin epsilon gets expressed in the primary cancer. P53 overexpression causes a decrease in the scores of HBB and HBA. In stage III there are decreases in the scores of HBB and HBA. P53 knockout causes increases in all proteins scores. Adding vitamin D or vitaminD+P53 overexpression seem to provoke unclear effects in these hemoglobolin subunits scores. Pre-malignant tissues may express hemoglobin subunits to levels which do not cause harm. However, raised expression or appearance of these proteins in the primary tumors may prompt survival and proliferation of tumor cells (5). However, overexpression of P53 may hinder cancer cell proliferation. In stage III there are decreases in the scores of HBB and HBA, perhaps this explains the efficacy of adjuvant chemotherapy at this stage (8).

Gastric cancer

We analyzed by proteomics the secretion of different hemoglobin by different gastric cancer cell lines derived from primary tumor tissues or lymph node metastases or stage IV tumors in cell culture. We found out that, HBG-2 is present in both primary cancer (SNU1) and lymph node (LN) metastases (HGC27) . From primary cancer to LN metastases, HBG-2 score raises considerably. HGC27 also shows the presence of HBB and HBA. Curiously, stage IV cells (SNU16) do not show any types of hemoglobins (9).

Prostate cancer

By proteomics analysis of cell CM precipitates, derived from cell culture of prostate cancer cells lines such as PC3, DU145, Vcap, and LNcap, we found that all these cell lines secrete HBB with a range of different scores, being that LNcap (lymph node metastasis) only secretes HBB and with a low score. Both PC3 (bone metastasis, negative to androgen receptor) and DU145 (brain metastasis) secrete HBA with a low score. Moreover, Vcap (bone metastasis, wild type (WT) androgen receptor), additionally expresses HBD with a level 2 score. Finally, PC3 also secretes hemoglobin gamma-2 (HG2) with a low score (5-8)

Lung cancer

We analyzed by proteomic analysis, the precipitates derived from CM of different lung cancer cell lines, all NSCLC and also derived from the culture of a bone metastases from lung cancer sample. In what concerns, adenocarcinomas, HBB and HBA scores raise from primary cancer to lymph node (LN) metastases. HBG2 may appear with a NS (non-significant) score in primary cancer, but becomes significant in LN metastases. In other NSLCs types, HBB score raises from primary cancer to LN metastases. Finally, bone metastases, show a spectacular raise on HBB and HBA, appears HBD with a very strong score, also HBG1 and HBE and HBZ (hemoglobin zeta) is non-significant (10).

Head and Neck cancer

We analyzed by proteomics the CM precipitates of two different samples of head and neck cancer samples culture, M2 and M9. We found that HBB and HBD are consistently found in both samples with significant Mascot scores (11).

Bone cancer

Cell lines such as 143B and SKES1 express HBB, HBA and HBG2, cell lines such as SAOS, additionally express HBD and U2OS does not express HBG2, but expresses a different HBD variant (E9PFT6) (12).

Children’s cancers

We analyzed by proteomic analysis, the precipitates derived from CM of IMR5, SKNBe2a and CCG9911 cell lines. All these cells lines express HBA and HBB. Only neuroblastoma cell lines express HBE. HBG2 is expressed in SKNBe2a and CCG9911 cell lines (13).

Acute leukemias

We found out that HBA, Mascot scores are raised in leukemia samples, while HBB Mascot scores are lower in leukemia samples. Also, HBD only is present in control samples (14).

For a summary, please, see Table 1.

References

1. Taylor A, Pollack, MA. Hemoglobin Level and Tumor Growth. Cancer Research (1942)

2. BMC Cancer. 2013 Jul 10;13:340. doi: 10.1186/1471-2407-13-340.Post-diagnosis hemoglobin change associates with overall survival of multiple malignancies - results from a 14-year hospital-based cohort of lung, breast, colorectal, and liver cancers. Wan S, Lai Y, Myers RE, Li B, Palazzo JP, Burkart AL, Chen G, Xing J, Yang H.

3.  Tucker R, Pedro A. Blood-derived non-extracellular vesicle proteins as potential biomarkers for the diagnosis of early ER+ breast cancer and detection of lymph node involvement. Version 3. F1000Res. 2018 Mar 6 [revised 2018 May 10];7:283. doi: 10.12688/f1000research.14129.3. eCollection 2018.

4. Beretov J, Wasinger VC,Millar EK, Schwartz P, Graham PH, Li Y.Proteomic Analysis of Urine to Identify Breast Cancer Biomarker Candidates Using a Label-Free LC-MS/MS Approach. PLoS One. 2015 Nov 6;10(11):e0141876. doi: 10.1371/journal.pone.0141876.

5. Non-conventional role of haemoglobin beta in breast malignancy. British Journal of Cancer (2017) 117, 994–1006 | doi: 10.1038/bjc.2017.247. Marco Ponzetti

6. Pedro A (2022). Hemoglobin as a biomarker for breast cancer ISSN 2753-8176 (online) (DOI: 10.13140/RG.2.2.25461.65764)

7. Pedro A (2022). Hemoglobin as a biomarker for melanoma - short report DOI: 10.13140/RG.2.2.32520.29445, ISSN 2753-8176 (online)

8. Pedro A (2022). Hemoglobin in colorectal cancer (DOI: 10.13140/RG.2.2.32515.99363, ISSN 2753-8176 (online))

9. Pedro A (2023).Hemoglobin in gastric cancer (ISSN 2753-8176 (online), DOI:10.1101/202291v1 , Small report

10. Pedro A (2023).Hemoglobin in Lung Cancer - Short report (ISSN 2753-8176 (online), DOI: 10.13140/RG.2.2.30186.59842

11.Pedro A (2023).Hemoglobin in Head and Neck cancer - Short report (ISSN 2753-8176 (online), DOI: 10.13140/RG.2.2.16792.67846

12. Pedro A (2023).Hemoglobin and osteoblasts markers in bone cancer, ISSN 2753-8176 (online), DOI: 10.13140/RG.2.2.14509.05606

13. Pedro A (2023).Hemoglobin and Children’s cancers - Brief Report, DOI: 10.13140/RG.2.2.21321.52325, ISSN 2753-8176 (online)

14. Pedro A (2023).Hemoglobin in Acute Leukemias - Brief report , ISSN 2753-8176 (online)

Amendments:

Curiously, as mentioned in the lung cancer and hemoglobin article, WI38 (3 month old female embryo lung tissue cell line) expresses hemoblobin beta (P68871),and hemoglobin alpha (P69905) with Mascot Scores of 348.33 and 969.56, respectively.