Immunohistochemical Study of the Over-Expressed Protein of P57-Gene Related withHPV31/33 Infections in a Group of Thyroid Carcinomatous Tissues

 

Nihaya Kadhim Salim¹, SaadHasan Mohammed Ali², Israa Mahdi Al-Sudani³

¹Ministry of Health, Baghdad, Iraq.

²Clinical Communicable Diseases Research Unit, College of Medicine, University of Baghdad.

³Ibn Sina University of Medical and Pharmaceutical Sciences, Iraq, Baghdad.

 

ABSTRACT:

Background: The incidence of thyroid cancer has increased during the last three decades. Many studies have been conducted to determine whether there is a link between viral infections and thyroid carcinogenesis. Human Papilloma viruses (HPV) are related to a variety of benign and malignant tumors. P57 is a member of the Cip/Kip family that binds and inhibits all cyclin/CDK complexes, resulting in cell-cycle arrest as well as cell growth. Objective: This retrospective study designed to investigate histopathological expression of thyroid cancer tissues in relation to the concordant HPV31/33 infection and p57 protein over-expressions. Patients and Methods: HPV31/33 DNA and P57-gene protein expression were evaluated in 116 thyroid tissues. The samples related to 36 patients with thyroid carcinoma, 40 thyroid adenoma, and 40 normal thyroid tissues. In situ hybridization (ISH) used to identify HPV31/33-DNA, whereas immunohistochemistry (IHC) used to assess P57-gene expression. Results: Regarding thyroid carcinoma group, HPV31/33DNA-ISH are responses observed in 77.8%, in 30 % of thyroid adenomas group, and in 22.5 %of normal thyroid tissues group. The difference between HPV31/33 in thyroid malignancies and the control group was statistically significant. The p57 protein detected in 75% thyroid cancer tissues and in 52.5 % adenoma thyroid tissues, whereas 35% of the investigated normal thyroid tissues showed IHC-reactions. The difference in the detected percentages of P57 protein was statistically significant in thyroid tissues between the study groupsin relations to the control group. Conclusions: The significant detection of HPV31/33 along with over-expressed P57-gene in thyroid cancer patients could support a possible role for both HPV 31/33 along with this p57 protein in thyroid pathogenesis and for carcinogenesis.

 

 

INTRODUCTION:

Human papilloma viruses are found to be completely species and tissue-specific viruses¹. To date, the family of Papillomaviridae comprises more than 450 different genotypes of human papillomavirus and over 200 animal papilloma viruses^2,3,4. The viruses are epithelio-tropic viruses infecting cutaneous tissues as well as mucosal membranes leading to asymptomatic infection, benign warts and malignancies⁵.

 

Infection with HPV affects a range of tissues among them, respiratory, oral cavity, ano-genital tract and urethral mucosal tissues¹. According to their oncogenic potential these viruses divided into high and low-risk types ⁶. The HPV16 and HPV18 globally recognized as a common cause of sexually transmitted viral infections that linked etiologically to >95% of cervical cancers ^{7, 8}.

 

Cancers, as non-communicable diseases, are now ranking as the leading cause of global deaths, expected as the single most important barrier globally for increasing life expectancy in the 21^st century (Ministry of Health\Environment^9,10. Over the past three decades, the incidence of thyroid cancer increased globally. Regarding the Eastern Mediterranean Region, the predicted cancer rates could be doubled by 2030¹¹. However, in selected high-income countries were in relation to the increased access to healthcare, improved diagnostic technology, and increased surveillance account for more than 60% of thyroid cancer diagnoses¹². The thyroid gland as a member of the endocrine system consists of two lobes (connected by the isthmus) as well as a third one, the pyramidal lobe, (found in 28- 55% of the population)¹³.

 

According to their morphology and biology, thyroid cancers are divided grossly into two groups; (1^st ) well differentiated thyroid cancers [slowly growing] papillary thyroid carcinoma (PTC)(accounts for approximately 85% of thyroid tumors) and the follicular thyroid carcinoma (FTC) (accounts for 5-15% of thyroid malignancies)^14,15, (2^nd)poorly differentiated thyroid cancers[rapidly growing and invasive] anaplastic thyroid cancers (ATC), (comprising a 1-2% of all diagnosed thyroid malignancies). Approximately 5% of thyroid malignancies originating from the parafollicular C cells diagnosed as medullary carcinoma¹⁴.

 

P57 belongs to the Cip/Kip family (one of two cyclin inhibitory families), it binds to and inhibits all cyclin/CDK complexes. Despite the apparent duplication of each Cip/Kip member (p21, p27, and p57), their siblings serve separate functions ^{16, 17}. P27 and p57 have similar structures, especially in their N-terminal regions, where a conserved domain has been identified that are needed for binding and inhibiting CDK complexes ^{18, 19}.

 

This study aimed to estimate the participation role of relevant high-oncogenic risk types of human papilloma virus (namely genotypes31/33) in archival tissues specimens from thyroid malignancies and benign tumors, as well as to examine the impact of the expression of p57 gene.

 

MATERIALS AND METHODS:

The study has selected 116 formalin fixed paraffin embedded thyroid tissues blocks; which included (36) malignant thyroid tumor divided into grade I,II & III, (40) thyroid adenoma tissues and 40 healthy thyroid tissues(used as control group).One section was specified to be mounted on ordinary glass slide and stained with hematoxylin and eosin, while other slides mounted on positive charged slides to be used for ISH for detection of HPV and for IHC to detection of p57 protein. In one hand, the detection of HPV31/33 by ISH kit (Zyto Vision GmbH. Fischkai, Bremerhaven. Germany) was performed on 4µm paraffin embedded tissue sections using digoxigenin-labeled oligo-nucleotides probes that target HPV31/33-DNA. Immunohistochemistry detection system (Abcam, England) used to demonstrate the P57 protein expression. Pearson Chi –square test by SPSS program (Version–27) & P value was considered significant when p ≤0.05.

 

RESULTS:

The age ofpatients with thyroid carcinoma ranged from 11 to 80 years, while those with thyroid adenoma from 10 to 67 years and those from where normal thyroid tissues obtained from 13 to 67 years. Statistically, non-significant differences noticed among research groups (P 0.840). The most affected age stratum among all the examined patients were those aged from 40-49 years as showed in table-1. Gender distribution detects are shown in table-1 where non-significant difference noticed.

 

Table 1: The characteristics of the study groups

	Thyroid Carcinoma	Thyroid Adenoma	Apparently Healthy Control	P value
-Age (years)	37.9±14.5 (11-80)	40.3±13.4 (10-67)	39.4±12.9 (13-67)	0.840
Gender
Male	8	8	9	0.957
Female	28	32	31
Stage
Stage I	29	-	-
Stage II	4	-	-
Stage III	3	-	-

-Data were presented as Mean±SD (Range)

 

The tumor stage of thirty-six patients with thyroid carcinoma investigated by a histopathologist, where 80.6% of total patients had stage I, 11.1% of patients had stage II, and 8.3% of patients had stage III, could be seen in table-1.

 

The HR-HPV31/33 DNA was identified in tissue blocks from 77.8% thyroid carcinomatous tissues, in 30% thyroid adenomatous tissues, and in22.5% thyroid control tissues group. The statistical differences between the percentages in these groups are highly significant (P 0.0001) and as indicated in(Table-2). Table-2 also shows the signal of scores and intensities positive HPV 31/33-CISH where 55.6% have high intensities and 55.6% with score3 in thyroid carcinoma tissues, 7.5% with high intensities and 17.5% have score3 in thyroid adenoma tissues while 17.5% have high intensities and 15% have score3 inapparently healthy control tissues. High statistical significant differences noticed among studied groups.

 

Table2: CISH detection of DNA HR-HPV (31/33) in the studied groups

CISH		Thyroid Carcinoma		Thyroid Adenoma		Apparently Healthy Control		P value
		No	%	No	%	No	%
HPV31,33	Positive	28	77.8	12	30.0	9	22.5	0.0001
	Negative	8	22.2	28	70.0	31	77.5
HPV31,33 Intensity	Low	3	8.3	6	15.0	-	-	0.0001
	Moderate	5	13.9	3	7.5	2	5.0
	High	20	55.6	3	7.5	7	17.5
HPV31,33 Score	1	2	5.6	1	2.5	1	2.5	0.0001
	2	6	16.7	4	10.0	2	5.0
	3	20	55.6	7	17.5	6	15.0

 

Table3: CISH detection of DNA HR-HPV (31/33) in the studied groups

CISH		Thyroid Carcinoma		Thyroid Adenoma		Apparently Healthy Control		P value
		No	%	No	%	No	%
HPV31,33	Positive	28	77.8	12	30.0	9	22.5	0.0001
	Negative	8	22.2	28	70.0	31	77.5
HPV31,33 Integrated pattern	<10	-	-	1	8.3	2	22.2	0.279
	10-19	1	3.6	-	-	2	22.2
	20-29	2	7.1	2	16.7	1	11.1
	30-39	2	7.1	1	8.3	-	-
	40-49	3	10.7	-	-	-	-
	50-59	2	7.1	-	-	-	-
	60-69	1	3.6	-	-	-	-
	70-79	1	3.6	1	8.3	1	11.1
	80-89	-	-	1	8.3	-	-
	=>90	16	57.1	6	50.0	3	33.3
	Mean±SD (Range)	71.8±30.3 (15-100)		66.8±35.8 (5-97)		47.0±43.5 (5-98)
HPV31,33 Diffused pattern	<10	14	50.0	5	41.7	3	33.3	0.110
	10-19	2	7.1	2	16.7	-	-
	20-29	-	-	-	-	1	11.1
	30-39	1	3.6	1	8.3	-	-
	40-49	1	3.6	-	-	-	-
	50-59	2	7.1	-	-	-	-
	60-69	4	14.3	-	-	-	-
	70-79	2	7.1	2	16.7	-	-
	80-89	2	7.1	1	8.3	2	22.2
	=>90	-	-	1	8.3	3	33.3
	Mean±SD (Range)	28.2±30.2 (2-85)		33.2±35.8 (3-95)		53.0±43.5 (2-95)

*Significant difference between percentages using Pearson Chi-square test (c2-test) at 0.05 level

 

Table-3 shows the physical status of the DNA-expressed patterns where the means of HPV31/33 DNA expression as an integrated form in thyroid carcinoma tissues were 71.8±30.3, in thyroid adenoma tissues, the mean of HPV31/33 DNA expression as an integrated form was 66.8±35.8, and the mean of HPV31/33 DNA expression as an integrated form of apparently healthy control tissues was 47.0±43.5, high significant differences among studied groups figure-1.

 

Figure 1: HR-HPV genotype 31/33 DNA physical patterns

In this work, the IHC method was employed to detect changes in p57-expression in the cells of the investigated thyroid tissues in the different study groups. Table-4 demonstrates that positive p57-IHC values obtained in 75% of thyroid carcinoma tissues, in 52.5% of thyroid adenoma tissues, and positive p57-IHC values were found in 35% of the apparently healthy control tissues with high-significant differences (p 0.002). Table-4, additionally, demonstrates high intensities as well as score3 in studied groups, 47.2% and 61.1%, respectively in thyroid carcinoma tissues, 40% and52.5%, respectively, of thyroid adenoma tissues, and 32.5%, 35%, respectively of normal thyroid control tissues. There were statistically significant differences between the studied groups.

 

 

Table 4: IHC-expression of p57 protein among studied groups

IHC		Thyroid Carcinoma		Thyroid Adenoma		Apparently Healthy Control		P value
		No	%	No	%	No	%
P57	Positive	27	75.0	21	52.5	14	35.0	0.002
	Negative	9	25.0	19	47.5	26	65.0
P57 Intensity	low	6	16.7	1	2.5	-	-	0.003
	moderate	4	11.1	4	10.0	1	2.5
	high	17	47.2	16	40.0	13	32.5
P57 Score	1	2	5.6	-	-	-	-	0.002
	2	3	8.3	-	-	-	-
	3	22	61.1	21	52.5	14	35.0

*Significant difference between percentages using Pearson Chi-square test (c2-test) at 0.05 level

 

Immunostaining for all the proteins was classified on the basis of the percentage of stained cells as follows: 0, negative; +1, ˂10 percent; +2,10 to 49 percent; +3, ≥50 percent (figure-2).

 

Figure2: Results of immunohistochemistry, in situ hybridization, and histology. (A) Hematoxylin and eosin staining of thyroid tissue; (B) HPV31/33-ISH displays primarily blue staining. HPV31/33-ISH positive result (C) displays primarily Red staining negative result (D) P57 immunostaining was found to be positive (dark brawn) (E) Immunohistochemistry of P57 yielded a negative result (purple hematoxylin stain).

 

DISCUSSION:

This study found a statistically highly significant difference in the detection rates of HPV 31/33 among the analyzed thyroid tumors and control groups by using the CISH technique in conjunction with the histopathological expressions of thyroid tumors (i.e. thyroid carcinoma and thyroid adenoma) and as compared to apparently healthy thyroid tissues. Mostafaeiet al., 2020¹⁵, revealed that papillary carcinoma (among thyroid tumor types) was the most commonly related to viral infections, where it is likely due to the fact that papillary cancer is the most frequent kind of thyroid carcinoma, accounting for nearly 85% of cases. As shown in table-3, HR-HPV31/33-DNA positive results were found in77.8% ofthyroid carcinomatous tissues,in thyroid adenomatous tissues 30% showed positive results,and 22.5% revealed positive HPV31/33results in control thyroid tissues with (P 0.0001).

 

In Iraq, Mohammed et al.²⁰, conducted a study in 2018 to identify the expressedhigh oncogenic risk HPV-18 antigen in thyroid tissues by using immunohistochemistry technique and found that 56.7% of patients had HPV-18 antigen, with 35% of benign thyroid tumors have HPV-18 positive-IHC tests while none of the healthy tissues in the control tissues having HPV-18 positive- IHC reactions. By the analogy of both HPV18 & HPV31/33 as high-oncogenic risk HPV genotypes, these findings are relatively consistent with the results of the present study, although those researchers studied the HPV 18- expressed antigens in these tissues while the present study, as a pioneer study in Iraq, searched for HPV 31/33 DNA localization where the differences in rates are referred for the difference in technique, viral genotype, and the physical states of the examined HPV genotype, as HPV-DNA viruses antigen expression.

 

In 2021, Dialameh, et al.²¹, (from Shiraz, Iran) considered their study to be the first in Iran, that link HPV to thyroid cancers (particularly papillary thyroid carcinoma) when they enrolled sample of 82 papillary thyroid carcinoma (PTC) and 77 benign thyroid nodules and used nested polymerase chain reaction technique to determine the presence of a broad spectrum of HPV genotypes and they found that 13.4% had positive-results for nPCR HPV of PTC and 3.8% had positive-results for nPCR HPV of benign thyroid nodules samples, while none of the neighboring normal tissues showed PCR positive results. In that work, Dialameh, et al. and associates highly urged that future studies in this field to be employ by different methods (other than PCR) for HPV-detection such as chromogenic in situ hybridization and immunohistochemistry. In both studies (the Dialameh’s study and the present study) it could be suggested that the infecting particles of HPV can possibly reached the thyroid gland via the bloodstream. Globally, the head and neck malignant tumors ranked the 6^th most common malignancies where more than 70% of them reported in third world countries²². Furthermore, several investigations for the identification of HPV in head and neck malignancies, such as Ali et al.²³ studies in Iraq in 2011 and 2017, reported that 3.2% and 10%, respectively, of positive CISH signals for HPV DNA in a group of apparently healthy oral and nasal control tissues. Another study in Iraq by Ali, et al.²³ in 2021 found that 20% of the positive CISH-HPV-DNA results in the adenoid hypertrophy group, while the control tissues revealed no test result. This research also revealed that the integrated form of HPV-DNA-CISH signal detection in adenoid hypertrophy tissues was noticed in 70% of the positive HPV results in the adenoid hypertrophy.

 

P57-IHC positive results there were high statistical significant differences among these thyroid tissue groups (p 0.002) as in Table-4. In analogy, the cancer development is the result of several combining and interacting variables involve multiple genes and processes. Recurrence and metastasis are serious issues that pose hazards to human health while biotechnology has progressed, more genes that may promote or prevent cancer development discovered. So as to explore the relationships of the genes to the cancer growth this is critical for predicting cancer prognosis as well as treatment efficacy^24,25,26. The decreased p57 expression was found to be linked to tumor aggressiveness and poor prognosis in a variety of cancers. However, mutations in the p57 gene (CDKN1C) are uncommon in malignancies^27,28.

 

Lee, et al. (2010)²⁹ revealed that because the cell cycle is controlled at the level of the nucleus, earlier investigations of cell-cycle regulators have neglected their cytoplasmic expression and focused primarily on their nuclear expression, suggesting that loss their function plays a role in malignant transformation. According to Lee et al. 2010, the upregulation of a tumor suppressor as a result of other molecular alterations that allow the cell to enter the cell cycle causes the cell to over express cytoplasmic p57. Upregulation of cytoplasmic p57 might indicate a cell's attempt to halt cell cycle growth and keep cell cycle control (i.e., a homeostatic feedback mechanism). Nuclear P57 expression was found in 57.1% of follicular adenoma cases, 65.6% of papillary thyroid carcinoma cases without lymph node metastasis, and 78.1% of papillary thyroid malignant tumors with lymph node metastasis. However, in the cytoplasm of tumor cells from the PTC group, there was some positive expression of p57. The sensitivity of immunostaining for cytoplasmic p57 was 43.75% in a research by Lee et al. 2010²⁷. According to Borrielloet al., (2011) ³⁰, three investigations on the level of p57 in thyroid tumors have been conducted. Ito et al. (2002) ³¹ examined the expression of p57 and found the protein was overexpressed in thyroid follicular adenomas and minimally in invasive thyroid follicular adenomas, while in an invasive follicular carcinomas showed a decrease in protein express. In poorly differentiated and undifferentiated carcinomas, p57 expression was reduced. In 2004, the same research group found that p57 protein was significantly reduced in more than 50% of the thyroid lymphoma cases. However, Melck et al. (2007)³² found no difference in p57-expression.

Oncoproteins of HPV may interact with proteins that regulated cell cycle like CDKs and CDK inhibitors. In cervical cancer, there is some evidence that HPV E7 can inhibit the action of a tumor suppressor called p21Cip1 (Cip/Kip family members p21, p27,and p57). HPV genotype 16-E7 inhibits p21's action on proliferating cell nuclear antigen (PCNA)-dependent DNA replication. Several investigations also revealed that inhibiting p63 expression in HPV31-positive keratinocytes lowered the expression of cyclins A, B, and E, as well as Cdk1, Cdc25c, and Cdk2^33,34.

 

CONCLUSIONS:

In conclusion, the considerable identification of HPV31/33 together with the over-expressed protein from the P57-gene in patients with thyroid cancer (and other studied series of thyroid adenoma as well as apparently healthy thyroid tissues) could point for a critical role (among other multi-factors in thyroid carcinogenesis) where along with the changes in the expression of CDK and CIKs (have long been thought to be a characteristic of cancer progression). Given the relevance of CIKs in thyroid tissues, not only management indicators but also treatment approaches to viral infections might be identified.

 

RECOMMENDATION:

We recommended other molecular research in the near future with large sample size and more clinicopathological features in different thyroid cancers to detect the presence of HR-HPV31/33-DNA and to assess the role of HPV high-risk genotypes in carcinogenesis (initiation and progression) of thyroid tumors. Study the role of p57 and its importance in the regulation of cell cycle and survival in benign and malignant tumors of the thyroid gland by using more developed laboratory materials and techniques to right predict diseases and choice the effective therapeutic patterns.

 

REFERENCES:

1.     Marimuthu M. Viswanathan T. Radha M. Suganya J. Computational Screening of the Phytocompounds from the Plant Ballota nigra Linn against the Human Papillomavirus (HPV) E6. Research Journal of Pharmacy and Technology 2017; 10(9): 3095-3097.  DOI: 10.5958/0974-360X.2017.00549.2

2.     Alameedy FMM. Diagnosis of Human Papillomavirus in Colorectal Cancer and Inhibition by Gamma Radiation. Research J. Pharm. and Tech. 2019; 12(5):2182-2184. DOI: 105958/0974-36x201900390.1

3.     Yi MS. Siang TC. Lwin S. Myint KT. Yee KT. Et al. Prevalence of Human Papilloma virus in women with Abnormal Cervical Smears from Sarawak, Malaysia. Research Journal of Pharmacy and Technology. 2021; 14(5):2729-4. DOI: 10.52711/0974-360X.2021.00481

4.     Santacroce L. Di Cosola M. Bottalico L. Topi S. Charitos IA. et al. Focus on HPV Infection and the Molecular Mechanisms of Oral Carcinogenesis. Viruses 2021; 13: 559. doi: 10.3390/v13040559.

5.     ParveenA, Jough SS. Study on the Cytotoxic Impacts of Thymol as the Segment of Trachyspermum ammi on Bosom Disease (MCF-7) Cells. Asian Journal of Research Chemistry. 2020; 13(5):327-333. DOI:10.5958/0974-4150.2020.00063.2

6.     Rani BSS. HPV Infection and Cervical Cancer. International Journal of Nursing Education and Research 2015; 3(2): 229-231.

7.     Mezher MN. Hashim AAA. Relationship of Human Papilloma Virus (HPV) and Epstein Barr Virus (EBV) with Prostate Cancer in AL-Najaf Governorate. Research Journal of Pharmacy and Technology 2017; 10(10): 3283-3288. DOI: 10.5958/0974-360X.2017.00582.0

8.     Nicole SL. Ito Y-TY. Jha S. High-Risk Human Papillomaviral Oncogenes E6 and E7 Target Key Cellular Pathways to Achieve Oncogenesis. International Journal of Molecular Science 2018; 19: 1706. doi: 10.3390/ijms19061706

9.     Ministry of Health/Environment, 2018. Iraqi Cancer Board. Annual Report Iraqi Cancer Registry 2016. P23

10.  Bray F. Ferlay J. Soerjomataram I. Siegel RL. Torre LA. Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 2018; 68: 394–424. doi: 10.3322/caac.21492

11.  WHO/ Cancer. Fact sheet, February 2018: http://www.who.int/mediacentre/factsheets/fs297/en/

12.  Singh ON, Iniguez-Ariza NM. Castro MR. Thyroid nodules: diagnostic evaluation based on thyroid cancer risk assessment. BMJ 2020; 368: l6670. doi: 10.1136/bmj.l6670

13.  De Sanctis V. Soliman AT. Di Maio S. Elsedfy H. Soliman NA. Elalaily R. Thyroid hemiagenesis from childhood to adulthood: review of  literature and personal experience. Pediatric Endocrinology Review 2016; 13(3): 612-619.

14.  Shakir H. Ausama A. Amber A. Saad H. Ahmed A. Molecular Localization of Human Papillomaviru-18 (HPV-18) in Tissues from Thyroid Carcinoma in Mid-Euphrate-Iraq. Advances in Natural and Applied Sciences 2015; 9(14): 25-31

15.  Mostafaei S. Keshavarz M. Sadri Nahand J. FarhadiHassankiadeh R. Moradinazar M. Nouri M. Moghoofei M. Viral Infections and Risk of Thyroid Cancer: A Systematic Review and Empirical Bayesian Meta-Analysis. Pathology - Research and Practice 2020; 2016(4): 152855. doi: 10.1016/j.prp.2020.152855

16.  Al-Ezzi BIH. Qasim BJ. Abid WB. Al-Ezzi MI. The Relationship between Clinic Pathological Characteristics and Follicular Thyroid Lesion. Research Journal of Pharmacy and Technology 2018; 11(2):741-744. DOI: 10.5958/0974-360X.2018.00139.7

17.  Hussein MJ. Al-Alwany SHM. Molecular Highlighting Analysis of Mutational CDK41, CDK6 and P27 in association with Human Mammary Tumor Virus Infection in Tissues from Iraqi Female with Ovarian Tumors. Research Journal of Pharmacy and Technology 2018; 11(10): 4699-4706. DOI:10.5958/0974-360X.2018.00858.2

18.  Russo GL. Stampone E. Cervellera C. Borriello A. Regulation of p27Kip1 and p57Kip2 Functions by Natural Polyphenols. Biomolecules 2020; 10: 1316. DOI: 10.3390/biom10091316

19.  Xie P. Wang H. Xie J. Huang Z. Chen S. et al. USP7 promotes proliferation of papillary thyroid carcinoma cells through TBX3-mediated p57KIP2 repression. Molecular and Cellular Endocrinology 2020; 518: 111037. doi: 10.1016/j.mce.2020.111037

20.  Mohammed SH. Alajeely AAA. AbdulAmeer AAAH. MohammedAli SH. Abed-Azuwaid AA.  Molecular Localization of Human Papillomaviru_18 (HPV_18) in Tissues from Thyroid Carcinoma in Mid-Euphrates. Journal of University of Babylon, Pure and Applied Sciences, Vol. (26),  No.(6).

21.  ArchinDialameh P. Saki F. Monabbati A. Dehghanian A. Valibeigi B. Soveid M. Detection of Human Papillomavirus in Papillary Thyroid Carcinoma and its Association with Tumor Staging and Pathologic Features. Iranian Journal of Medical Sciences 2021; 46(4): 256–262. doi: 10.30476/ijms.2020.83135.1191

22.  Al-Alwany S. Kadhim A. Al_Saffar H. Al-jebory AK. et al. Molecular Detection of Cytomegalovirus, CDK2 and P27 in Tissues from Patients with Thyroid Carcinoma.Journal of Global Pharma Technology 2017; 10 (9): 19-29.

23.  Ali SHM. Al-Hijazi AY. Khashman BM. P53-tumor suppressor gene overexpression in human papilloma virus-infected patients with oral squamous cell carcinoma. Journal of Baghdad College of Dentistry 2011; 23(sp. issue): 70-76.

24.  Ali SHM, Hussien IA, Hussein TA, Kamal MS. Nuclear in Situ Hybridization of High –Risk and Low-Risk Human Papilloma Virus DNA in Tissues from a Group of Iraqi Patients with Sino Nasal and Nasopharyngeal Benign and Malignant Tumors. Journal of Global Pharma Technology 2017; 10(9):362-370.

25.  Ali SH. Mohammed KI. Ali WM. Al-Fakhar SA. Al-alwany SH. Mousa J.M. In Situ Hybridization for Molecular Detection of Human Papilloma Viral 6 / 11 DNA in Adenoctomized Tissues from A group of Iraqi Pediatric Patients. Baghdad Science Journal 2022; 19: 0026. DOI: 0.21123/bsj.2022.19.1.0026

26.  Ru Y. Chen XJ. Zhao ZW. Zhang PF. Feng SH. et al. CyclinD1 and p57kip2 as biomarkers in differentiation, metastasis and prognosis of gastric cardia adenocarcinoma. Oncotarget 2017; 8(43): 73860–73870. doi: 10.18632/oncotarget.18008

27.  Stampone E. Caldarelli I. Zullo A. Bencivenga D. Mancini FP. et al. Genetic and epigenetic control of CDKN1C expression: importance in cell commitment and differentiation. Tissue Homeost, and human diseases. International Journal of Molecular Sciences 2018; 19(4): 1055. doi: 10.3390/ijms19041055

28.  Creff J. Besson A. Functional Versatility of the CDK Inhibitor p57Kip2. Front. Cell Dev. Biol 2020; 8: 584590. doi: 10.3389/fcell.2020.584590

29.  Lee SH. Lee JK. Jin SM. Lee KC. Sohn JH. et al. Expression of cell-cycle regulators (cyclin D1, cyclin E, p27kip1, p57kip2) in papillary thyroid carcinoma. Otolaryngology–Head and Neck Surgery 2010; 142(3): 332–337. doi: 10.1016/j.otohns.2009.10.050

30.  Borriello A. Caldarelli I. Bencivenga D. Criscuolo M. Cucciolla V. et al. p57(Kip2) and cancer: time for a critical appraisal. Molecular Cancer Research 2011; 9(10): 1269–1284. doi: 10.1158/1541-7786.MCR-11-0220.

31.  Ito Y. Yoshida H. Matsuzuka F. Matsuura N. Nakamura Y. et al. Expression of the components of the Cip/Kip family in malignant lymphoma of the thyroid. Pathobiology 2004; 71: 164–70. DOI: 10.1159/000076472

32.  Melck A Masoudi H. Griffith OL. Rajput A. Wilkins G. et al. Cell cycle regulators show diagnostic and prognostic utility for differentiated thyroid cancer. Ann Surg Oncol 2007; 14(12): 3403–3411. doi: 10.1245/s10434-007-9572-8

33.  Tavakolian S. Goudarzi H. Faghihloo E. Cyclin-dependent kinases and CDK inhibitors in virus-associated cancers. Infectious Agents and Cancer 202; 15: 27. DOI: 10.1186/s13027-020-00295-7

34.  Yadav K. Singh D. Singh MR. Development and Characterization of Corticosteroid loaded Lipid carrier system for Psoriasis. Research Journal of Pharmacy and Technology 2021; 14(2): 966-970. DOI:10.5958/0974-360X.2021.00172.4

 

 

 

Accepted on 20.04.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(11):5011-5016.