Lung cancer is considered as one of the most common cause of death. The major reason, why lung tumor is so common is due to the much feared HPV and also smoking. The major association between lung tumor and HPV infection has been testified, some people think that it was due to geographical and racial diversify. The main aim behind this research is to discover the existence of human papilloma virus in lung cancer patients by making use of immunohistochemcial method. In this particular study, a total of thirty five (35) lung cancer patients were studied, out of which 30 were male and 5 were female patients. The ages of patients ranged from 36-96 years, with 30 of them being smokers. HPV expressed in lung cancer tissue, were especially common in smoker patients.The study proved that human papilloma virus played a substantial role as a major risk factor in lung cancer.
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Lung cancer is one of the main reason behind deaths of both genders all across the world and number of these death are increasing day by day(Brambilla et al., 2001). This trend of increasing rate of death is predominantly due to the fact that patients with lung cancer are discovered at a later stage of cancer development. Mortality rate can be reduced to a great extent, If early detection of lung cancer can be developed somehow. Proper treatment procedure of lung cancer can only be conceivable, if early detection of cancerous cell can be achieved somehow. This is a very serious issue as the lung is a possible place of carcinoma and malignancy, and it could even further turn into a more fatal disease (Mittal KR et.al., 1993). Lung cancer particularly grows in two separate compartments of the lung, one is the peripheral (respiratory) part of the lung and the other is the central (transporting air) portion of the lung.
There are at least four major categories of cancer which have been indentified, including small cell cancer, non-small cell cancer, large cell cancer and many others types which are not that frequent. Lengthy exposure of the epithelium results in lining of lungs with numerous carcinogenic elements, primarily due to smoking of cigarette, which can eventually result in a frequent genetic variations of the cells, which would ultimately result in numerous concerns related to health. These various variations could result in morphological concerns, including development and propagation of tumor cells (Cina SJ et.al., 2001). Subsequently, lung tumor could also possibly turn into a alveoli or major bronchus. Carcinoma is generally positioned in central part of the lung, whereas large cell cancer are characteristically positioned in the peripheral part of the lung (Nillson K et.al., 2006). HPV is involved within pathogenesis in certain individual’s carcinomas like carcinomas of cervix and oropharynx (Syrjänen KJ et.al., 2000). Human papilloma virus are usually linked by the development of cervical tumors. Human papilloma virus of type 16 and type 18 kinds are repeatedly related with sever inside the cell layers, in addition to malignant neoplasm (Syrjänen KJ et.al., 1979). DNA of Human papilloma virus was discovered, after a large number of biopsies within diverse separated suitcases of lung carcinoma (Nadji SA et.al., 2007).
Twenty different type of cases with lung tumor are assimilated in this study in order to make a method of immunohistochemical to categorize Human Papilloma Virus in Lung Cancer tissue removed from a patient. All These patients used in study had thoracotomy along with bronchoscopic biopsy as well. All of these thirty five (35) lung cancer patients blocks had been inserted with paraffin wax and also fixed with formalin. First of all, I will categorize patients on the basis of age group, on the basis of gender and on the basis of smokers and non smokers. 30 male and 5 female patients, where involved in this study. The ages of pateints were ranging from 36-96 years with average age 63.55 years. 31 patients out of these were smokers. The ages of these lung cancer patients are characterized to certain age groups: from 30-40 there is only one case, which is a male patient. From 40-50 years, there are two male cases, from 51-60 years, there is one female case, nine male cases and two female cases, from 61-70 there were five male and two female case. From 71-80 years, there were nine male cases and one female cases, while age group greater than eighty years, had only one female case. Total sum of male Patients are thirty (30) and female patients were five (5). The number of patient who smoked are around thirty one, including 25 male and 6 female patients. During this study, there were around five cases of adenocarcinoma, twenty five cases of squamous cell carcinoma, and five cases were small cell carcinoma. During this research, Formalin fixed inserted chunk of benign skin was used as a control for diagnosis of Human Papilloma Virus in lung cancer tissues.
In this study, The immunohistochemistry staining method used were labeled streptavidin biotin technique which were practically designed for Human Papilloma Virus discoloration. The paraffin embedded and formalin fixed tissue subjected to microtome section to sections in thickness. All of these sections were dehydrated in a oven for 60 minutes at 60 °C temperature. After that dewaxation was done by dipping of each slides in xylene baths for 5 minutes, after that rehydration was done by repeated dilutions of alcohol, in two time alteration of 100% ethanol, 90% ethanol and 80% ethanol for three minute, for each of these cases. For further rehydration it was placed in phosphate buffer saline clean bath for room temperature for almost thirty minutes. After that, Slide are cleaned by means of deionized Water, After that they were transferred in a microwave oven in citrate buffer two times for duration of five minutes. Extra solution is get rid of with awareness and by means of a quick movement. Put 100 µl Anti-Human Papilloma (HPV) which is monoclonal antibody solution, coat the tissue slice. Incubation was done for almost sixty minutes at 37 °C temperature in a humidify hall. After that, 100 µl Biotinylated link Antibody was placed in all slides, to coat the tissue slice. Incubation for almost thirty minutes was done again at room temperature. Then all slides were put in a 100 µl Streptavidin. Incubation for almost twenty minutes in humid chamber was done at room temperature. Then put adequate drops of DAB+ and incubate for five minutes. Washing the slides gently for five minutes in water would do the required job. One of the slide was treated with primary antibody to use it as a positive control, second slide was not incubated with primary antibody, so that it could be used as a negative control.
The typical factor for positive immuno-reactions is dark brown in color. Each type of cancer was specified with a certain score based on the amount of the nuclear staining (Han CP et.al 2008). The immuno-reactive score was then calculated by taking the product of the intensity of positive scores of stained cells and extent of them as well. The minimum score was set at 0, while maximum score was set at 12 (Cuello, 1993). Variation of Human papilloma virus among age groups, gender, smoking status, cell types, were calculated by making use of Pearson’s Chi-square test. Research showed that HPV immuno=expression was reported in seven out of thirty five cases of lung cancer and wenty six cases were negative. This immuno-detection rate looks positively well connected to patients who are indulged in smoking. The immunoexpression of Human papilloma virus is also well related to age groups in a big way.
The results of this study have confirmed that eight out of thirty five cases of lung carcinoma are express Human papilloma virus revealed by using immuohistochemical cytoplasmic staining, which seem to be brown in color while staining. Out of those twenty seven case, seven cases were of squamous cell carcinoma cases, one case was of adenocarcinoma. whereas zero case out of six cases, were diagnosed with small cell carcinoma. This lung cancer showed almost 20 percent positively for Human papilloma virus. The immunon histochemical examination of the result shows that HPV is prevalent in seven out of thirty two cases, who are also smokers and one out of eight cases for nonsmokers. Research showed that there was no correlation between detection of Human papilloma virus and smoking history of the patient, But generally the rate of Human papilloma virus infection is lower in nonsmokers than smokers.
Our study nearly agreed with the study conducted by (Kato et al., 2012) but it is not in line with the study conducted by (Ya-Wen Cheng et.al., 2007) that showed that Human papilloma virus was found out in large numbers for nonsmoker patients than smokers. immunoexpression of HPV revealed that not any of female cases were exposed with HPV but eight out of thirty male were exposed with Human papilloma virus. This difference of stats of HPV prevalence among gender group were not so significant, which is in accordance to the study performed by (Yang Fei1et.al., 2010 Human papilloma virus was found out more in case of male patients, higher than female with p value greater than 0.05. whereas, research by (Cheng YW et al., 2007) showed that women were HPV expressed more than their male counterparts. The immuno expression of Human papilloma virus, in relation to different age groups, revealed that no HPV exposure was shown in age group between thirty and forty years, only one case between age 41 and 50 out of three cases, four cases between age 51 and 60 out of thirteen cases, two cases between age group of 61 and 70 out of ten cases, one case between age group 71 and 80 years out of tweleve cases, and finally no case were reported in age group greater than 80 years. The difference among these ages groups shows the effect of HPV exposure in a very different way. The immuno histochemical method for detection of this issue seems to be slightly higher with age group between 51 and 60 years which is also in correlation with the study performed by (Cheng YW et al., 2007)
The study concluded that the prevalence of Human papilloma virus has been found nearly 20% among the lung cancer cases, mostly in squamous cell carcinomas. Small cell carcinoma tissues did not demonstrate any sign of Human papilloma virus when they were studied using immuno histochemical method. Detection of Human papilloma virus in lung carcinoma can easily be achived by using a well correlated method of immunohistocemical technique. Detection and treatment of these symptoms is one thing but preventation is far better than cure. One has to realize about the threats of lung cancer and how it can affect a person both physically and mentally. Study confirmed that smoking is one of the possible reason for lung cancer but still rate of active smokers are increasing day by day. A person has to realize that despite the invention of all these modern medical procedures, a person can still suffer, if the lung cancer is not diagnosed earlier. So prevention is always better than cure.
Ullman R, Bongiovanni M, Halbweld I, Petzmann S, GoggKammerer M, Sapino A, Papotti M, Bussolati G, Popper HH.(2003) Bronchiolar columnar cell dysplasia – genetic analysis of anovelpreneoplastic lesion of peripheral lung. Virchows Arch, ; 442: 429-436
Cina SJ, Lancaster-Weiss KJ, Lecksell K, Epstein JI. (2001). Correlation of Ki-67 and p53 with the new World Health Organization/International Society of Urogical Pathology Classification System for UrothelialNeoplasia. Arch Pathol Lab Med,; 125: 646- 651
Brambilla E, Travis WD, Colby TV, Corrin B, Shimosato Y, (2001). The new World Health Organisation classification of lungtumor. EurRespir J 18: 1059-68
Mittal KR, Demopoulos RI, Goswami S. (1993) Proliferating cellnuclear antigen (cyclin) expression in normal and abnormalcervical squamous epithelia. Am J SurgPathol,;17: 117-22
Jensen-Taubman S, Steinberg SM, Linnoila RI. (1998) Bronchiolization of the alveoli in lung cancer: pathology, patterns ofdifferentiation and oncogene expression. Int J Cancer,;75: 489-496
Nillson K, Landberg G. (2006). Subcellular localization, modification and protein complex formation of the cdk-inhibitor p16 in Rb-functional and Rb-inactivated tumor cells. Int J Cancer,; 118: 1120-1125
Syrjänen KJ and Syrjänen S: (2000). Papillomavirus infection in human pathology. John Wiley & Sons, New York, pp1-10; 355-378.
Nadji SA, Mokhtari-Azad T, Mahmoodi M, et al: (2007). Relationship between lung
cancer and human papillomavirus in north of Iran,Mazandaran province. Cancer Lett
Syrjänen KJ (1980). Epithelial lesions suggestive of a condylomatousorigin found closely
associated with invasive bronchial squamouscell carcinomas. Respiration 40: 150-160,
Kamoi S, AlJuboury MI, Akin MR, et al. (2000) Immunohistochemical staining in the
distinction between primary endometrial and endocervical adenocarcinomas: another
viewpoint. Int J Gynecol Pathol;21:217–223.
Syrjänen KJ. (1979) Condylomatous changes in neoplastic bronchial epithelium. Report of a case. Respiration 38: 299-304 .