PTEN and PI3K Mutation Markers and Expression of CD68 and IL-6 Inflammatory Markers in Endometrioid and Clear Cell Ovarian Carcinoma with Underlying Endometriosis

Arsad N; Abd Aziz NH; Tizen Laim NMS; Md Zin RR; Abdul Hafizz AMH; SHAFIEE MN

Original Article Published on 14 Mar, 2024

PTEN and PI3K Mutation Markers and Expression of CD68 and IL-6 Inflammatory Markers in Endometrioid and Clear Cell Ovarian Carcinoma with Underlying Endometriosis

Mutation and Inflammatory Markers in Ovarian Cancer

Vol. 19 No. 1 : 2024 (257-270)

Arsad N

Abd Aziz NH

Tizen Laim NMS

Md Zin RR

Abdul Hafizz AMH

SHAFIEE MN

Abstract

The objective of this research endeavour was to ascertain whether there existed a distinction in the protein expression levels of phosphatase and tensin homolog (PTEN), phosphatidylinositol-4,5-biphosphate3-kinase (PI3K), macrophage (CD68), and interleukin-6 (IL-6) among women who had been diagnosed as having ovarian carcinoma (endometrioid and clear cell subtypes) underlying endometriosis. Tissue microarray (TMA) slides were fabricated using donor’s formalin-fixed paraffin-embedded (FFPE) blocks that comprised specimens of ovarian carcinomas tissues with the subtypes of endometrioid and clear cell that were retrieved for a duration of nine years. A total of 19 FFPE blocks represented 19 cases of carcinoma of the ovary, which were categorised into two groups: ovarian cancer with endometriosis (EAOC, n=10) and ovarian cancer without endometriosis (n=9). All of these protein expressions were analysed using immunohistochemistry (IHC) technique. Following that, comparisons were made regarding the clinical and pathological characteristics of the ovarian carcinomas, as well as any discernible variations in expression levels between the two groups. In EAOC, PTEN expression was lower (88.9% vs. 100%, P=0.47) compared to those without endometriosis. Our findings revealed an increase of PI3K and IL-6 in EAOC than those devoid of endometriosis; 80% vs. 77.8%, P=1.00 and 70% vs 11.11%, P=0.35, respectively. In contrast, CD68 was significantly lower in endometriosis-associated ovarian cancer compared to those without endometriosis (40% vs. 66.67%, P=0.16). In conclusion, there were no significant difference in the mutational and inflammatory marker modifications between clear cell carcinoma and endometrioid adenocarcinoma of ovary with endometriosis.

Keywords : CD68, clear cell adenocarcinoma, endometrioid adenocarcinoma, endometriosis, interleukin-6, ovarian carcinoma, PI3K, PTEN,

Abstrak

Objektif penyelidikan ini adalah untuk memastikan sama ada terdapat perbezaan dalam tahap ekspresi protein fosfatase dan tensin homolog (PTEN), fosfatidilinositol-4,5-bifosfat3-kinase (PI3K), makrofaj (CD68) dan interleukin-6 (IL-6) di kalangan wanita yang telah didiagnosis menghidap karsinoma ovari (subjenis endometrioid dan sel jernih) yang mendasari endometriosis. Slaid tisu mikrorai (TMA) telah disediakan dengan menggunakan blok parafin terfiksasi formalin (FFPE) penderma yang mengandungi spesimen tisu karsinoma ovari dengan subtaip endometrioid dan karsinoma sel jernih pada ovari yang telah diambil untuk tempoh sembilan tahun. Sebanyak 19 blok FFPE terdiri daripada 19 kes kansasinoma ovari, yang dikategorikan kepada dua kumpulan iaitu kanser ovari dengan endometriosis (EAOC, n=10) dan kanser ovari tanpa endometriosis (n=9). Kesemua ekspresi protein ini telah dianalisis menggunakan teknik imunohistokimia (IHC). Setelah itu, perbandingan berkaitan dengan ciri-ciri klinikal dan patologi karsinoma ovari, serta sebarang variasi yang boleh dilihat dalam tahap ekspresi antara kedua-dua kumpulan telah dilakukan. Dalam EAOC, ekspresi PTEN adalah lebih rendah (88.9% berbanding 100%, P=0.47) berbanding mereka yang tidak mempunyai endometriosis. Penemuan kami mendedahkan peningkatan PI3K dan IL-6 dalam EAOC daripada mereka yang tidak mempunyai endometriosis; masing-masing terdapat 80% berbanding 77.8%, P=1.00 dan 70% berbanding 11.11%, P=0.35. Sebaliknya, CD68 adalah jauh lebih rendah dalam kanser ovari yang berkaitan dengan endometriosis berbanding mereka yang tidak mempunyai endometriosis (40% berbanding 66.67%, P=0.16). Kesimpulannya, terdapat perbezaan ketara dalam pengubahsuaian penanda mutasi dan keradangan antara karsinoma sel jernih dan adenokarsinoma endometrioid ovari dengan endometriosis.

Kata Kunci : Adenokarsinoma endometrioid, adenokarsinoma sel jelas, CD68, endometriosis, interleukin-6, karsinoma ovari, PI3K, PTEN,

Correspondance Address

Professor Dr. Mohamad Nasir Shafiee. Department of Obstetrics and Gynaecology, Faculty of Medicine, Universiti Kebangsaan Malaysia (UKM), Jalan Ya’acob Latiff, Bandar Tun Razak, Cheras, Kuala Lumpur, Malaysia. Tel: +603-91455949 Email: nasirshafiee@ukm.edu.my

References

Abdul Karim, A.K., Abd Aziz, N.H., Md Zin, R.R., Mohd Mokhtar, N., Shafiee, M.N. 2020. The effect of surgical intervention of endometriosis to CA-125 and pain. Malays J Med Sci 27(6): 7-14

Ankasha, S.J., Shafiee, M.N., Wahab, N.A., Ali, R.A.R., Mokhtar, N.M. 2018. Post-transcriptional regulation of microRNAs in cancer: From prediction to validation. Oncol Rev 12(1): 344.

Azam, I.N.A., Wahab, N.A., Mokhtar, M.H., Shafiee, M.N., Mokhtar, N.M. 2022. Roles of microRNAs in regulating apoptosis in the pathogenesis of Endometriosis. Life 12(9): 1321.

Azizah, A.M., Nor Saleha, I.T., Noor Hashimah, A., Asmah, Z.A., Mastulu, W. 2016. Malaysian national cancer registry report 2007–2011: Malaysia cancer statistics, data and figure. Putrajaya: National Cancer Registry.

Cheaib, B., Auguste, A., Leary, A. 2015. The PI3K/Akt/mTOR pathway in ovarian cancer: Therapeutic opportunities and challenges. Chin J Cancer 34(1): 4-16.

Dewhurst, J. 2012. Dewhurst’s textbook of obstetrics and gynaecology. New Jersey: John Wiley & Sons.

Hao, T., Ge, A., Dong, W., Han, F., & Song, H. 2017. Clinical significances of PTEN and HER-2 protein expression in endometriosis associated ovarian carcinomas and their correlations. Biomed Res 28(22): 9734-8.

Isobe, A., Sawada, K., Kinose, Y., Ohyagi-Hara, C., Nakatsuka, E., Makino, H., Ogura, T., Mizuno, T., Suzuki, N., Morii, E. 2015. Interleukin 6 receptor is an independent prognostic factor and a potential therapeutic target of ovarian cancer. PloS One 10(2): e0118080.

Kalamanathan, S., Bates, V., Lord, R., Green, J. 2011. The mutational profile of sporadic epithelial ovarian carcinoma. Anticancer Res 31(8): 2661-8.

Krawczyk, N., Banys-Paluchowski, M., Schmidt, D., Ulrich, U., Fehm, T. 2016. Endometriosis-associated malignancy. Geburtshilfe und Frauenheilkunde 76(02): 176-81.

Kriplani, D., Patel, M.M. 2013. Immunohistochemistry: A diagnostic aid in differentiating primary epithelial ovarian tumors and tumors metastatic to the ovary. South Asian J Cancer 2(4): 254-8.

Kuo, K.T., Mao, T.L., Jones, S., Veras, E., Ayhan, A., Wang, T.L., Glas, R., Slamon, D., Velculescu, V.E., Kuman, R.J. 2009. Frequent activating mutations of PIK3CA in ovarian clear cell carcinoma. Am J Pathol 174(5): 1597-601.

Lane, D., Matte, I., Rancourt, C., Piché, A. 2011. Prognostic significance of IL-6 and IL-8 ascites levels in ovarian cancer patients. BMC Cancer 11(1): 1-6.

Li, S., Fu, X., Wu, T., Yang, L., Hu, C., Wu, R. 2017. Role of interleukin-6 and its receptor in endometriosis. Med Sci Monit 23: 3801-7.

Makker, A., Goel, M.M., Das, V., Agarwal, A. 2012. PI3K-Akt-mTOR and MAPK signaling pathways in polycystic ovarian syndrome, uterine leiomyomas and endometriosis: An update. Gynecol Endocrinol 28(3): 175-81.

Musa, F., Schneider, R. 2015. Targeting the PI3K/AKT/mTOR pathway in ovarian cancer. Transl Cancer Res 4(1): 97-106.

Nezhat, F.R., Apostol, R., Nezhat, C., Pejovic, T. 2015. New insights in the pathophysiology of ovarian cancer and implications for screening and prevention. Am J Obstet Gynecol 213(3): 262-7.

Orezzoli, J., Russell, A., Oliva, E., Del Carmen, M., Eichhorn, J., Fuller, A. 2008. Prognostic implication of endometriosis in clear cell carcinoma of the ovary. Gynecol Oncol 110(3): 336-44.

Pearce, C.L., Templeman, C., Rossing, M.A., Lee, A., Near, A.M., Webb, P.M., Nagle, C.M., Doherty, J.A., Cushing-Haugen, K.L., Wicklund, K.G. et al. 2012. Association between endometriosis and risk of histological subtypes of ovarian cancer: A pooled analysis of case–control studies. Lancet Oncol 13(4): 385-94.

Prat, J. 2012. Ovarian carcinomas: Five distinct diseases with different origins, genetic alterations, and clinicopathological features. Virchows Archiv 460(3): 237-49.

Rao, S.R., Sy-Cherng, L.W., Hafizz, A.M.H.A., Mamat Yusof, M.N., Shafiee, M.N. 2023. Sexual functioning and marital satisfaction among endometriosis patients in Malaysia: A cross-sectional study. Front Psychol 14: 1224995.

Romero, I., Bast Jr, R.C. 2012. Minireview: Human ovarian cancer: Biology, current management, and paths to personalizing therapy. Endocrinology 153(4): 1593-602.

Sato, N., Tsunoda, H., Nishida, M., Morishita, Y., Takimoto, Y., Kubo, T., Noguchi, M. 2000. Loss of heterozygosity on 10q23.3 and mutation of the tumor suppressor gene PTEN in benign endometrial cyst of the ovary: Possible sequence progression from benign endometrial cyst to endometrioid carcinoma and clear cell carcinoma of the ovary. Cancer Res 60(24): 7052-6.

Shafiee, M.N., Lim, W.K., Shi, C.P.S., Yasin, I.A.M., Azemi, A.F., Zakaria, M.L., Hafizz, A.M.H.A., Mustangin, M., Kampan, N.C., Aziz, N.H.A., Zain, R.R.M. 2021. PTEN protein expression has role in predicting disease-free-interval in endometrioid endometrial carcinoma. Horm Mol Biol Clin Investig 42(4): 403-10.

Somigliana, E., Parazzini, F., Stoppelli, S., Giambattista, E., Vercellini, P. 2006. Association between endometriosis and cancer: Acomprehensive review and a critical analysis of clinical and epidemiological evidence. Gynecol Oncol 101(2): 331-41.

Vestergaard, A.L., Thorup, K., Knudsen, U.B., Munk, T., Rosbach, H., Poulsen, J.B., Guldberg, P., Martensen, P.M. 2011. Oncogenic events associated with endometrial and ovarian cancers are rare in endometriosis. Mol Hum Reprod 17(12): 758-61.

Wang, L., Wang, C., Jin, S., Qu, D., Ying, H. 2015. Expression of NF-κB and PTEN in primary epithelial ovarian carcinoma and the correlation with chemoresistance. Int J Clin Exp Pathol 8(9): 10953.

Yamamoto, S., Tsuda, H., Takano, M., Iwaya, K., Tamai, S., Matsubara, O. 2011. PIK3CA mutation is an early event in the development of endometriosis-associated ovarian clear cell adenocarcinoma. J Pathol 225(2): 189-94.

Yousefi, H., Momeny, M., Ghaffari, S.H., Parsanejad, N., Poursheikhani, A., Javadikooshesh, S., Zarrinrad, G., Esmaeili, F., Alishahi, Z., Sabourinejad, Z. 2019. IL-6/IL-6R pathway is a therapeutic target in chemoresistant ovarian cancer. Tumori J 105(1): 84-91.

Correspondance Address

Abdul Karim, A.K., Abd Aziz, N.H., Md Zin, R.R., Mohd Mokhtar, N., Shafiee, M.N. 2020. The effect of surgical intervention of endometriosis to CA-125 and pain. Malays J Med Sci 27(6): 7-14

Ankasha, S.J., Shafiee, M.N., Wahab, N.A., Ali, R.A.R., Mokhtar, N.M. 2018. Post-transcriptional regulation of microRNAs in cancer: From prediction to validation. Oncol Rev 12(1): 344.

Azam, I.N.A., Wahab, N.A., Mokhtar, M.H., Shafiee, M.N., Mokhtar, N.M. 2022. Roles of microRNAs in regulating apoptosis in the pathogenesis of Endometriosis. Life 12(9): 1321.

Azizah, A.M., Nor Saleha, I.T., Noor Hashimah, A., Asmah, Z.A., Mastulu, W. 2016. Malaysian national cancer registry report 2007–2011: Malaysia cancer statistics, data and figure. Putrajaya: National Cancer Registry.

Cheaib, B., Auguste, A., Leary, A. 2015. The PI3K/Akt/mTOR pathway in ovarian cancer: Therapeutic opportunities and challenges. Chin J Cancer 34(1): 4-16.

Dewhurst, J. 2012. Dewhurst’s textbook of obstetrics and gynaecology. New Jersey: John Wiley & Sons.

Hao, T., Ge, A., Dong, W., Han, F., & Song, H. 2017. Clinical significances of PTEN and HER-2 protein expression in endometriosis associated ovarian carcinomas and their correlations. Biomed Res 28(22): 9734-8.

Isobe, A., Sawada, K., Kinose, Y., Ohyagi-Hara, C., Nakatsuka, E., Makino, H., Ogura, T., Mizuno, T., Suzuki, N., Morii, E. 2015. Interleukin 6 receptor is an independent prognostic factor and a potential therapeutic target of ovarian cancer. PloS One 10(2): e0118080.

Kalamanathan, S., Bates, V., Lord, R., Green, J. 2011. The mutational profile of sporadic epithelial ovarian carcinoma. Anticancer Res 31(8): 2661-8.

Krawczyk, N., Banys-Paluchowski, M., Schmidt, D., Ulrich, U., Fehm, T. 2016. Endometriosis-associated malignancy. Geburtshilfe und Frauenheilkunde 76(02): 176-81.

Kriplani, D., Patel, M.M. 2013. Immunohistochemistry: A diagnostic aid in differentiating primary epithelial ovarian tumors and tumors metastatic to the ovary. South Asian J Cancer 2(4): 254-8.

Kuo, K.T., Mao, T.L., Jones, S., Veras, E., Ayhan, A., Wang, T.L., Glas, R., Slamon, D., Velculescu, V.E., Kuman, R.J. 2009. Frequent activating mutations of PIK3CA in ovarian clear cell carcinoma. Am J Pathol 174(5): 1597-601.

Lane, D., Matte, I., Rancourt, C., Piché, A. 2011. Prognostic significance of IL-6 and IL-8 ascites levels in ovarian cancer patients. BMC Cancer 11(1): 1-6.

Li, S., Fu, X., Wu, T., Yang, L., Hu, C., Wu, R. 2017. Role of interleukin-6 and its receptor in endometriosis. Med Sci Monit 23: 3801-7.

Makker, A., Goel, M.M., Das, V., Agarwal, A. 2012. PI3K-Akt-mTOR and MAPK signaling pathways in polycystic ovarian syndrome, uterine leiomyomas and endometriosis: An update. Gynecol Endocrinol 28(3): 175-81.

Musa, F., Schneider, R. 2015. Targeting the PI3K/AKT/mTOR pathway in ovarian cancer. Transl Cancer Res 4(1): 97-106.

Nezhat, F.R., Apostol, R., Nezhat, C., Pejovic, T. 2015. New insights in the pathophysiology of ovarian cancer and implications for screening and prevention. Am J Obstet Gynecol 213(3): 262-7.

Orezzoli, J., Russell, A., Oliva, E., Del Carmen, M., Eichhorn, J., Fuller, A. 2008. Prognostic implication of endometriosis in clear cell carcinoma of the ovary. Gynecol Oncol 110(3): 336-44.

Pearce, C.L., Templeman, C., Rossing, M.A., Lee, A., Near, A.M., Webb, P.M., Nagle, C.M., Doherty, J.A., Cushing-Haugen, K.L., Wicklund, K.G. et al. 2012. Association between endometriosis and risk of histological subtypes of ovarian cancer: A pooled analysis of case–control studies. Lancet Oncol 13(4): 385-94.

Prat, J. 2012. Ovarian carcinomas: Five distinct diseases with different origins, genetic alterations, and clinicopathological features. Virchows Archiv 460(3): 237-49.

Rao, S.R., Sy-Cherng, L.W., Hafizz, A.M.H.A., Mamat Yusof, M.N., Shafiee, M.N. 2023. Sexual functioning and marital satisfaction among endometriosis patients in Malaysia: A cross-sectional study. Front Psychol 14: 1224995.

Romero, I., Bast Jr, R.C. 2012. Minireview: Human ovarian cancer: Biology, current management, and paths to personalizing therapy. Endocrinology 153(4): 1593-602.

Sato, N., Tsunoda, H., Nishida, M., Morishita, Y., Takimoto, Y., Kubo, T., Noguchi, M. 2000. Loss of heterozygosity on 10q23.3 and mutation of the tumor suppressor gene PTEN in benign endometrial cyst of the ovary: Possible sequence progression from benign endometrial cyst to endometrioid carcinoma and clear cell carcinoma of the ovary. Cancer Res 60(24): 7052-6.

Shafiee, M.N., Lim, W.K., Shi, C.P.S., Yasin, I.A.M., Azemi, A.F., Zakaria, M.L., Hafizz, A.M.H.A., Mustangin, M., Kampan, N.C., Aziz, N.H.A., Zain, R.R.M. 2021. PTEN protein expression has role in predicting disease-free-interval in endometrioid endometrial carcinoma. Horm Mol Biol Clin Investig 42(4): 403-10.

Somigliana, E., Parazzini, F., Stoppelli, S., Giambattista, E., Vercellini, P. 2006. Association between endometriosis and cancer: Acomprehensive review and a critical analysis of clinical and epidemiological evidence. Gynecol Oncol 101(2): 331-41.

Vestergaard, A.L., Thorup, K., Knudsen, U.B., Munk, T., Rosbach, H., Poulsen, J.B., Guldberg, P., Martensen, P.M. 2011. Oncogenic events associated with endometrial and ovarian cancers are rare in endometriosis. Mol Hum Reprod 17(12): 758-61.

Wang, L., Wang, C., Jin, S., Qu, D., Ying, H. 2015. Expression of NF-κB and PTEN in primary epithelial ovarian carcinoma and the correlation with chemoresistance. Int J Clin Exp Pathol 8(9): 10953.

Yamamoto, S., Tsuda, H., Takano, M., Iwaya, K., Tamai, S., Matsubara, O. 2011. PIK3CA mutation is an early event in the development of endometriosis-associated ovarian clear cell adenocarcinoma. J Pathol 225(2): 189-94.

Yousefi, H., Momeny, M., Ghaffari, S.H., Parsanejad, N., Poursheikhani, A., Javadikooshesh, S., Zarrinrad, G., Esmaeili, F., Alishahi, Z., Sabourinejad, Z. 2019. IL-6/IL-6R pathway is a therapeutic target in chemoresistant ovarian cancer. Tumori J 105(1): 84-91.