Journal of Cancer Research and Therapeutics

ORIGINAL ARTICLE
Year
: 2020  |  Volume : 16  |  Issue : 4  |  Page : 874--877

Heme oxygenase-1 in osteosarcoma


Simmi Kharb, M Halder, ZS Kundu 
 Department of Biochemistry and Orthopedics, Pt. B. D. S. PGIMS, Rohtak, Haryana, India

Correspondence Address:
Simmi Kharb
#1396, Sector-1, Rohtak, Haryana
India

Abstract

Aim of Study: The present study was planned to analyze serum heme oxygenase-1 levels in osteosarcoma patients. Materials and Methods: Twenty five histopathologically confirmed cases of osteosarcoma localized without metastasis of all the ages attending the Orthopedic Clinics were included in the study group and twenty five patients having musculoskeletal pain (age and sex matched) served as control. Five ml of venous blood was collected aseptically from antecubital vein and serum was be separated by centrifugation and analyzed the same day. Routine biochemistry investigations were performed as per standard enzymatic methods by autoanalyzer. Serum Heme oxygenase-1 was analyzed by enzyme-linked immunosorbent assay. Results: In osteosarcoma patients, serum HO-1 levels were increased as compared to patients having musculoskeletal pain (P < 0.05). Workers have found that HO-1 induction in prostate cancer cell lines (PC3) cells restored the proliferation of osteoblasts, which was inhibited during co-culture with parental prostate cancer cell line PC3 cells. However, no concrete data are available on blood levels of HO in osteosarcoma. Major role of HO-1 is the protection against oxidative injury, additionally, it regulates cell proliferation, modulates inflammatory response and facilitates angiogenesis. Conclusion: Findings of the present study suggests that pharmacological agents that regulate HO activity or HO-1 gene silencing may become powerful tools for preventing the onset or progression of various cancers and sensitize them to anticancer therapies.



How to cite this article:
Kharb S, Halder M, Kundu Z S. Heme oxygenase-1 in osteosarcoma.J Can Res Ther 2020;16:874-877


How to cite this URL:
Kharb S, Halder M, Kundu Z S. Heme oxygenase-1 in osteosarcoma. J Can Res Ther [serial online] 2020 [cited 2020 Oct 25 ];16:874-877
Available from: https://www.cancerjournal.net/text.asp?2020/16/4/874/237385


Full Text



 Introduction



Osteosarcoma is the most common primary malignant bone tumor, predominantly affecting children and adolescents. The role of heme oxygenase-1 (HO-1) in bone pathophysiology is not completely understood, and the status of HO-1 is not clear in osteosarcoma. Osteosarcoma is the most common bone tumor seen in the pediatric and adolescent age group. Osteosarcoma patient presents with painful swelling of bones.[1] Males are affected 1.5–2 times more than females with predilection for metaphysis of the long bone of lower limb.[1] In most patients, the etiology of osteosarcoma remains obscure. Several markers for diagnosis and prognosis have been proposed in osteosarcoma, namely, vascular endothelial growth factor, bone alkaline phosphatase, osteocalcin, survivin, and epidermal growth factor receptor ErbB2.[2]

HO catalyzes the oxidation of heme to generate carbon monoxide (CO), biliverdin, and iron. Three distinct mammalian HO isoforms (HO-1, HO-2, and HO-3) have been identified, and HO-1 is inducible isoform. HO-1 is expressed at low levels in most tissues under normal basal conditions and has been implicated in several physiological functions throughout the body, including the control of vascular tone, cytoprotective function, and regulation of the inflammatory and apoptotic cascades as well as contributing to the antioxidant capacity in several organ systems.[3]

The extent to which HO-1 per se mediates disease states such as diabetes and hypoxia remains to be fully clarified. Furthermore, expression of HO-1 is usually increased in tumors, compared with surrounding healthy tissues. This has been shown in lymphosarcoma, adenocarcinoma, hepatoma, glioblastoma, melanoma, prostate cancers, Kaposi sarcoma, squamous carcinoma, pancreatic cancer, and brain tumors.[4]

HO-1 is a proangiogenic mediator, which augments vascularization of tumors and increases the metastatic potential of cancer cells. Angiogenesis is critical not only for tumor growth but also for metastasis.[5]

The role of HO-1 in bone pathophysiology is not completely understood. HO has been reported to be have a negative regulatory role on osteoclast formation and bone resorption. Researchers have found that HO-1 induction in PC3 cells restored the proliferation of osteoblasts, which was inhibited during coculture with parental prostate cancer cell line PC3 cells.[6] However, no concrete data are available on blood levels of HO-1 in osteosarcoma.

The status of HO-1 is not clear in osteosarcoma, and hence, the present study was planned to analyze serum HO-1 levels in osteosarcoma patients and compare them with controls (patients with musculoskeletal pain).

 Materials and Methods



The present study was conducted in the Department of Biochemistry in collaboration with Department of Orthopedics, Pt. B. D. Sharma, PGIMS, Rohtak. Fifty patients were selected for the study and were divided into 2 groups, namely, Group I (n = 25, study): histopathologically confirmed cases of osteosarcoma (before starting treatment) and Group II (n = 25, control): age- and sex-matched patients with musculoskeletal pain (before giving any pain medication). Five-milliliter venous samples were drawn aseptically from antecubital vein and serum separated by centrifugation and analyzed the same day. Routine biochemistry investigations were performed as per standard enzymatic methods by autoanalyzer. Serum HO-1 was estimated by double-antibody sandwich enzyme-linked immunosorbent assay kits (Elabscience kit).[4]

IBM SPSS Statistics (Statistical Package for the Social Sciences) ver. 23 (IBM, Elabscience, Huston, Texas, USA) was used for statistical analyses. Results were expressed as mean ± standard error (standard deviation), and unpaired “t-” test and Pearson correlation test were applied. Data were considered to be statistically significant if P < 0.05.

 Results



Various biochemical parameters are shown in [Table 1]. Majority of osteosarcoma cases (n = 19, 76%) were in the age group of 10–20 years while 6 (24%) were in the age group of 21–30 years. Fourteen (56%) of the musculoskeletal pain cases were in the age group of 10–20 years while the remaining 11 (44%) were in the age group of 21–30 years [Table 1]. Twelve (48%) of the osteosarcoma patients were male and 13 were female (52%). In musculoskeletal patients, there were 8 males (32%) and 17 females (68%).{Table 1}

Hemoglobin levels were lower and serum bilirubin levels were higher in osteosarcoma cases as compared to controls. Serum calcium and alkaline phosphatase levels were significantly raised with osteosarcoma (Group I) as compared to controls (Group II) [Table 1]. Serum phosphorus levels were lower in the Group I as compared to Group II (P < 0.05). Serum alkaline phosphatase levels showed positive correlation with serum calcium levels in controls (P > 0.05) which got inversed in osteosarcoma cases (r = 0.132 vs. r = –0.131; P > 0.05). Serum alkaline phosphate levels showed negative correlation with serum phosphorus levels in controls (P > 0.05) which got inversed in osteosarcoma cases (r = –0.081 vs. r = 0.032; P > 0.05).

In osteosarcoma patients, serum HO-1 levels were increased as compared to controls with musculoskeletal pain (3.23 ± 0.31 [1.53] vs. 2.21 ± 0.27 [1.33] ng/ml; P < 0.05, [Figure 1]). Serum HO-1 levels showed a positive correlation with hemoglobin levels in controls (P > 0.05) which got inversed in osteosarcoma cases (r = 0.285 vs. r = –0.206; P > 0.05). Serum HO-1 levels showed significant negative correlation with bilirubin levels in controls (P < 0.001) which got inversed in osteosarcoma cases (r = –0.028 vs. r = 0.947; P > 0.05). Serum alkaline phosphatase levels showed negative relationship with serum HO-1 levels in both osteosarcoma and controls (r = –0.051 and r = –0.075; P > 0.05 at both levels). Serum HO-1 levels showed positive correlation with serum calcium levels in controls (P > 0.05) which got inversed in osteosarcoma cases (r = 0.0156 vs. r = –0.025; P > 0.05). Serum HO-1 levels showed positive correlation with serum phosphorus levels in controls (P > 0.05) which got inversed in osteosarcoma cases (r = 0.318 vs. r = –0.045; P > 0.05).{Figure 1}

 Discussion



Osteosarcoma is grouped as bone-forming tumors in the present study, and serum calcium levels were higher and serum phosphorus levels were lower in osteosarcoma cases as compared to controls in the present study [Table 1]. Furthermore, serum alkaline phosphatase levels were significantly raised in osteosarcoma cases as compared with controls (Group II, P < 0.001) [Table 1]. Elevated alkaline phosphate levels represent the increased osteoblastic activity.

HO-1 catabolizes heme into CO, biliverdin (which is rapidly converted to bilirubin), and free iron. HO-1 possesses important antioxidant and anti-inflammatory functions through antioxidant activities of biliverdin, bilirubin, and anti-inflammatory action of CO.[7]

Recent reports indicate that HO-1 is expressed in bone cells and may be involved in the regulation of bone homeostasis.[2],[3],[4],[5] In the present study, in osteosarcoma cases, the serum HO-1 levels were increased as compared to musculoskeletal pain controls (P < 0.05).

To the best of our knowledge, no report is available in literature where serum HO-1 levels have been reported in osteosarcoma. Researchers have found that HO-1 induction in PC3 cells restored the proliferation of osteoblasts, which was inhibited during coculture with parental prostate cancer cell line PC3 cells.[6]

Zwerina et al. reported that upregulation of HO-1 inhibits differentiation of osteoclasts from osteoclast precursors.[2] It has also been reported that HO-1 is induced in osteoblasts in weight-bearing bone of the rat by mechanical loading, indicating that it may participate the metabolism of osteoblasts.[7] HO-1 inhibits osteoblastic apoptosis induced by tumor necrosis factor α through generating CO.[8] and HO-1 also downregulates the production of cytokines by osteoblast.[9] However, the effects of HO-1 on osteoblastic differentiation are still not fully clarified.

In the present study, hemoglobin levels were lower and serum bilirubin levels were higher in osteosarcoma cases [Table 1]. In the present study, serum HO-1 levels showed a significant positive correlation with hemoglobin levels in Group II that turned to significant negative correlation in osteosarcoma group (r = –0.206, P < 0.05 at both levels). There was a negative correlation between HO-1 and serum bilirubin levels. This correlation became highly significant positive in osteosarcoma patients (r = –0.028, P < 0.05; r = 0.094, P < 0.001).

HO-1 downstream metabolites, bilirubin, and CO have been reported to have a role in the maturation of osteoblasts.[10] It has also been reported that bilirubin dose dependently inhibited the mineral deposition and differentiation markers of osteoblasts, suggesting that bilirubin may be partially responsible for the inhibitory effects of HO-1 on the osteoblast differentiation.[11] Findings of high serum HO-1 and high serum bilirubin levels in the osteosarcoma cases in the present study suggest the protective role of HO-1 and bilirubin in osteosarcoma.

Studies have shown that elevated HO activity could confer antioxidative protection to cells and tissues[4],[12] and elevation in serum bilirubin concentration with a significant and marked reduction in coronary heart disease.[5],[13],[14],[15] There is evidence that biliverdin/bilirubin can moderate some of HO-1 antiatherogenic effects,[6],[7],[16],[17] and bilirubin exerts anti-inflammatory and antiproliferative actions.[7],[16]

Lin et al. reported that bilirubin and CO have a role in the maturation of osteoblasts and bilirubin dose dependently inhibited the mineral deposition and differentiation markers of osteoblasts.[10] These findings suggested that bilirubin may be partially responsible for the inhibitory effects of HO-1 on the osteoblast differentiation.

However, no concrete data are available on blood levels of HO in osteosarcoma. On reviewing the literature, no study was available where HO-1 levels have been correlated with other biochemical parameters in osteosarcoma. Furthermore, no correlation between HO-1 with calcium and alkaline phosphatase has been reported.

Findings of the present study suggest that pharmacological agents that regulate HO activity or HO-1 gene silencing may become powerful tools for preventing the onset or progression of various cancers and sensitize them to anticancer therapies. Novel therapeutic modalities based on modifying the response to HO-1, modulation of redox species production, signaling, and metabolism and/or modulation of cellular antioxidant response may be effective in treating osteosarcoma in future.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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