IMMUNO HISTOCHEMISTRY OF STEM CELL MARKER CD 133 AND SMOKING IN LUNG CANCER PATIENTS-A CORRELATIVE STUDYHTML Full Text
IMMUNO HISTOCHEMISTRY OF STEM CELL MARKER CD 133 AND SMOKING IN LUNG CANCER PATIENTS-A CORRELATIVE STUDY
Neema Tiwari, Nishi Tandon *, A. N. Srivastava and Tanya Tripathi
Department of Pathology, Eras Lucknow Medical College, Lucknow - 226003, Uttar Pradesh, India.
ABSTRACT: Introduction: In India, lung carcinoma is the 5th most common tumor and 2nd most common tumor in males as per the ICMR [Indian Council of Medical Research] registry of 2002. In our Indian set up, smoking is still a leading cause of lung cancer and related morbidity and mortality. The role of many cancer stem cell markers like CD133, ALDH1, etc., is being investigated in the origin and prognosis of lung cancer. Cancer stem cells (CSCs) have gained increasing attention recently in cancer research. CSCs have the ability to generate new tumors through their stem cell properties, essentially self-renewal potential and differentiation into multiple cell lineages. Aim: we planned a study correlating Cancer stem cell marker CD133 with smoking in newly diagnosed lung cancer patients. Material and Methods: Lung biopsies of a total of 55 patients visiting Era’s Lucknow Medical College and hospital and King George Medical University, suspected to be suffering from lung cancer, were taken in this pilot study for a period of 18 months. Lung tissue fixed in 10% formalin processed in paraffin blocks, used for staining, Sigma for Haematoxylin and Eosin staining, and Protein Tech for Immunohistochemistry. A detailed history of tobacco exposure and smoking was taken, and the smoking index was calculated by using pack years. Result: No significant association was seen in the presence of CD133 and SI. However, an increasing trend was seen with CD133 presence and smoking. Conclusion: With a larger sample size, CD133 presence might correlate with exposure to smoking and hence impact the prognosis and origin of lung cancer.
Stem cell marker CD 133, Lung cancer
INTRODUCTION: Lung cancer is a very serious problem of the Indian subcontinent, especially in the lower socioeconomic subgroups. In India, lung carcinoma is the 5th most common tumor and the 2nd most common tumor in the males as per the ICMR [Indian Council of Medical Research] registry of 2002. It accounts for 6.9% of new cancer cases detected each year 1.
The absolute and relative frequency of lung cancer has risen dramatically over the decade. An example of the increasing frequency of lung cancers is that in the 19th century, the age-adjusted death rate from lung cancer was similar to that of pancreatic cancer; however, an increase has been seen since then in deaths caused by Lung carcinoma leading to increased age-adjusted death rates. In 1985 Lung carcinoma became the leading cause of cancer deaths in women and now causes approximately twice as many deaths as breast cancer. Of late, it has been seen that Lung cancer deaths are declining in men, and the death rate in women has plateaued secondary to decreases in smoking 2.
Exposure to biogas and chulha smoke, asbestosis, passive smoking are some other causes of lung cancer.
This is the WHO data and, in our Indian set up, smoking is still a leading cause of lung cancer and related morbidity and mortality. Cancer stem cell markers are being investigated not only in lung cancer but also in other varieties like colorectal cancers, breast cancers, and pancreatic cancers 3. Commonly being studied are CD133, ALDHI, ABPG2 etc. Hence, we planned a study correlating Cancer stem cell marker CD133 with smoking in newly diagnosed lung cancer patients.
MATERIALS AND METHODS: A total of 55 patients visiting Era’s Lucknow Medical College and hospital and King George Medical University, suspected to be suffering from lung cancer, were taken in this pilot study for a period of 18 months November 2014-April 2016. Patient consent and ethical clearance were taken before the study was conducted.
Patients with any co-morbidity like TB, fungal infections, or endocrine diseases were excluded. Lung tissue fixed in 10% formalin processed in paraffin blocks, used for staining, sigma for haematoxylin and eosin staining, and protein tech for immunohistochemistry. A detailed history of tobacco exposure and smoking was taken, and the smoking index was calculated by using pack years.
Statistical Analysis: The statistical analysis was done using SPSS (Statistical Package for Social Sciences) Version 15.0 statistical Analysis Software. The Mean, standard deviation students t test ANOVA Kruskall Wallis tools were applied for statistical analysis 4.
TABLE 1: DEMOGRAPHIC PROFILE OF THE CASES (n=55)
|Mean Age ± SD (Range) in years||53.51±10.76
FIG. 1: cd133 POSITIVITY IN LUNG CANCER ON 40x
TABLE 2: EXPOSURE TO DIFFERENT RISK FACTORS AND PATTERN OF EXPOSURE
|S. no.||Characteristic (in 55 cases)||Statistic|
|Biomass + Smoking||2 (3.6%)|
|2||Chewing tobacco/Gutka||8 (14.5%)|
|3||Mean Duration of smoking ± SD (Range) (n=43)||18.58±8.28
|4||Mean SI/FQ ±SD (Range) (n=43)||25.47±13.09
FIG. 2: THOUGH MEAN SFI OF THOSE WITH MARKER EXPRESSION WAS HIGHER AS COMPARED TO THOSE HAVING NO MARKER EXPRESSION, HOWEVER, THE ASSOCIATION WAS NOT SIGNIFICANT STATISTICALLY (P>0.05). Red-with marker expression, Green-Without marker expression
TABLE 3: ASSOCIATION BETWEEN SFI AND CD133/ALDH1 EXPRESSION (n=43)
|S. no.||Marker||SFI||Statistical Significance|
|Cases with marker expression||Cases without marker expression||‘t’||‘p’|
No significant association was seen in the presence of CD133 and SI
FIG. 3: HIGH POWER VIEW OF SQUAMOUS CELL CARCINOMA LUNG
FIG. 4: HIGH POWER VIEW OF A H&E SLIDE OF POORLY DIFFERENTIATED CARCINOMA LUNG
DISCUSSION: Lung cancer remains the leading cause of cancer-related mortality in the world and the most frequently diagnosed cancer worldwide, with non-small-cell lung cancer (NSCLC) accounting for about 80-85% of all lung cancer cases 5. There is a growing body of evidence that cancer stem cells (CSCs) represent a rare population of exclusively tumorigenic cells responsible for tumor initiation, progression, metastasis, and recurrence 6, 7. Therefore, a better understanding of the biology of CSCs is providing opportunities for improved cancer detection and therapy in the future. Various markers like aldh1, cd44, etc., have been proposed to define stem cell populations in distinct solid tumor types. Expression of the cell surface molecule CD133 is a well-accepted marker for lung CSCs.
Smoking is the most well-known etiological factor implicated in carcinogenesis. Both active and passive smoking or secondary smoking in the form of a non-smoker inhaling cigarette smoke exhaled by a smoker causes lung cancer. There are multiple genetic and epigenetic abnormalities associated with the pathogenesis of lung cancer 8. These abnormalities may result in the activation of oncogenes and inactivation of tumor-suppressor genes.
Chronic inflammation, which is known to promote cancer, may result both from smoking and from genetic abnormalities. These mediators, in turn, may be responsible for increased macrophage recruitment, delayed neutrophil clearance, and an increase in reactive oxygen species (ROS) 9. Thus, the pulmonary environment presents a unique milieu in which lung carcinogenesis proceeds in complicity with the host cellular network. This altered pulmonary microenvironment paves the way for both epithelial-mesenchymal transition (EMT) and the destruction of the specific host cell-mediated immune responses 10.
We in this study aimed at associating the expression of CSC CD133 with Smoking index [SI] in freshly diagnosed cases of lung cancer.
Statistical analysis was done using the chi-square test, Students T-test, with a comparison between parameters done using ANOVA and Kruskall Wallis H test. On studying the demographic data of the cases, we found that the age of patients under study ranged from 35 to 72 years. The majority of patients were aged above 50 years (54.5%), with the mean age of patients being 53.51±10.76 years. In the studies reviewed, the mean cut off age of patients taken was 40 years 117, 118, 119, while another study put the cut off age as 56 years 11.
Studying the sex distribution and exposure to risk factors of patients under study, we saw that majority of cases were males  (76.4%) while there were 13 (23.6%) females with the male to female ratio of patients being 3.23:1. Our finding was supported by another study done by Mohan A. et al., where they saw that the ratio of men to women was 7.4:1 12. In another study the male to female ratio was found to be increased (3.58:1), with 78 % of the total population being males 13.
On eliciting the clinical history, we saw that there were 42 (76.4%) smokers, 11 (20%) had exposure to biomass smoke, and 2 (3.6%) had exposure to biomass smoke as well as cigarette/bidi smoke. A total of 8 (14.5%) patients had an additional habit of tobacco/gutka (chewable tobacco products) chewing. The mean duration of smoking was 18.58 ± 8.28 years, and the SI/FQ index ranged from 10 to 60, with a mean value of 25.47 ± 13.09. These findings matched with another study where 73.8% were found to be smokers 14. Another study done on 255 patients saw 48% of patients having a significant smoking history 15. Smoking has been seen to be an important risk factor for the development of lung carcinoma and is well documented in literature 16, 17.
On studying the association between SFI and CD133 expression though the mean SFI of those with marker expression was higher as compared to those having no marker expression; however, the association was not significant statistically for either of two markers (p>0.05). While studying the association between CD133 marker expression score and SFI, in CD133 expression, with an increasing score, an increasing trend of mean SFI values was observed; however, it was not significant statistically (p=0.277) Higher expression of CD 133 with smoking has been found in some studies 18, 19. Biomass exposure is a phenomenon prevalent in the developing world like India, and our attempt was to highlight the importance of biomass exposure with conventional bidi smoking as a cause of lung cancer. The major limitations of this study were the small sample size, and absence of patient follow up, which prevent us from forming a definitive impression about the prognostic effect of these markers.
CONCLUSION: Not many studies have been done correlating the risk factor status with the CSC CD133 marker expression. Our study is a pilot study, so more data and a larger sample size are needed to get at a final definitive impression.
ACKNOWLEDGEMENT: I would like to thank the department of pulmonary medicine, Eras Lucknow medical college, Dept. of pulmonary medicine King George Medical University, Lucknow, Dept of CTVS, KGMU, Lucknow for their contribution and support. I would also like to thank my faculty in the department of pathology Eras Lucknow medical college and hospital.
CONFLICTS OF INTEREST: None declared
IEC Approval No.: ELMC/EC/R_Cell/2014/205
- Behera D: Epidemiology of lung cancer-Global and Indian perspective. JIACM 2012; 13(2): 131-7.
- Feng W: Identification of Human Lung Cancer Stem Cell Markers. Abstract: Research grant Program winning 2010.
- Salama R, Tang J and Gadgeel SM: Lung cancer stem cells: current progress and future perspectives. Jour of Stem Cell Research and Therapy 2012; S7: 007.
- Mydral G, Lambe M and Hilerdal G: Effect of delay on prognosis in patients with non small cell Lung cancer. Thorax 2004; 59: 45-49.
- Hosni HN, Daoud SA and Bassam AM: Immunohistochemical study of stem cell marker ALDH1 and BRCA1 in Breast Cancer. Academic Journal of Cancer Research 2014; 7(1): 01-07.
- Hanahan D and Weinberg RA: Hallmarks of cancer: the next generation. Cell 2011; 144(5): 646-74.
- Relation T, Dominici M and Horwitz EM: Concise Review: An (Im) Penetrable Shield: How the Tumor Microenvironment Protects Cancer Stem Cells. Stem Cells 2017; 35: 1123-30.
- Merkle FT, Ghosh S and Kamitaki N: Human pluripotent stem cells recurrently acquire and expand dominant negative P53 mutations. Nature 2017; 545: 229-33.
- Pawelek JM: Fusion of bone marrow-derived cells with cancer cells: metastasis as a secondary disease in cancer. Chin J Cancer 2014; 33: 133-9.
- Tomasetti C, Li L and Vogelstein B: Stem cell divisions, somatic mutations, cancer etiology, and cancer prevention. Science 2017; 355: 1330-4.
- Noronha V, Dikshit R and Raut N: Epidemiology of lung cancer in India: Focus on the differences between non-smokers and smokers: A single-centre experience. Ind J of Can 2012; 49(1): 74-81.
- Mohan A, Latifi AN and Guleria R: Increasing incidence of adenocarcinoma lung in India: Following the global trend? Ind J of Can 2015; 53(1): 92-95
- Deonarain MP, Kousparou CA and Epenetos AA: Antibodies targeting cancer stem cells: a new paradigm in immunotherapy? MAbs 2009; 1: 12-25.
- Roudi R, Korourian A, Shariftabrizi A and Madjd Z: Differential expression of cancer stem cell markers ALDH1 and CD133 in various lung cancer subtypes. Plos One 2015; 294-302.
- Prasetyanti PR and Medema JP: Intra-tumor heterogeneity from a cancer stem cell perspective. Mol Cancer 2017; 16: 41.
- Goldstraw P, Crowley J, Chansky K, Giroux DJ, Groome PA, Rami-Porta R, Postmus PE, Rusch V and Sobin L; International association for the study of lung cancer international staging committee; participating institutions. The IASLC Lung Cancer Staging Project: proposals for the revision of the TNM stage groupings in the forthcoming (seventh) edition of the TNM Classification of malignant tumours. J Thorac Oncol 2007; 2: 706-14.
- Noguchi M, Shimosato Y, Mills S(Ed.).Srenberg’s Diagnostic Surgical Pathology Virginia,Wolters Kluwer/Lippincott Williams and Wilkins 2015; Vol 1. 5th edition, 1053-97.
- Torre LA, Bray F and Siegel RL: Global cancer statistics, 2012. CA Cancer J Clin 2015; 65: 87-108.
- MacDonagh L, Gray SG and Breen E: Lung cancer stem cells: The root of resistance. Can Let 2016; 372: 147-56.
- Lindström A, Midtbö K and Arnesson LG: Fusion between M2-macrophages and cancer cells results in a subpopulation of radio resistant cells with enhanced DNA repair capacity. Oncotarget 2017; 8: 51370-86.
- Hardavella G, George R and Sethi T: Lung cancer stem cells-characteristics, phenotype. Trans Lung Cancer Res 2016; 5: 272-9.
How to cite this article:
Tiwari N, Tandon N, Srivastava AN and Tripathi T: Immuno histochemistry of stem cell marker CD 133 and smoking in lung cancer patients-a correlative study. Int J Pharm Sci & Res 2021; 12(2): 1251-55. doi: 10.13040/IJPSR.0975-8232.12(2).1251-55.
All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
N. Tiwari, N. Tandon *, A. N. Srivastava and T. Tripathi
Department of Pathology, Eras Lucknow Medical College, Lucknow, Uttar Pradesh, India.
28 February 2020
13 May 2020
21 May 2020
01 February 2021