zerumbone and Uterine-Cervical-Dysplasia

Cervical cancer is the second most common cause of cancer death in women. We have demonstrated previously that zerumbone (ZER) has an anti-cancer effect towards human cervical cancer cells (HeLa).. Anti-cancer properties of ZER were investigated using female Balb/c mice exposed prenatally to diethylstilboestrol. Female offspring have been treated with ZER (4, 8 and 16 mg/kg), normal saline and cisplatin (10mg/kg; positive control). The anti-cancer properties of ZER were evaluated using histopathology, TdT-mediated dUTP nick end labeling (TUNEL) Assay and immunohistochemical staining of Bcl-2-associated X protein (Bax), a key protein in mitochondrial pathway of apoptosis. In addition, laser capture microdissection microscopy isolated RNA was amplified using reverse transcriptase polymerase chain reaction (RT-PCR) based on the specific primer of B-cell lymphoma 2 (Bcl-2).. Treatment with ZER resulted (P<0.05, chi(2) statistics) in the regression of cervical intraepithelial neoplasia (CIN) resembling cisplatin effect (10mg/kg). TUNEL micrographs showed the absence of apoptosis in cancerous tissues treated with normal saline compared to ZER and cisplatin where abundant apoptotic cells were noticed. A post hoc analysis showed a significant (P<0.01) difference in mean percentage of apoptosis between normal saline treatment (0%), ZER (15.7%) and cisplatin (21.7%). Immunohistochemical staining of Bax protein revealed that ZER modulates the expression of this apoptosis marker. Administration of ZER has also modulated the expression of Bcl-2 gene.. These findings showed that ZER induces apoptosis efficiently in cervical tissues from female Balb/c mice treated prenatally with diethylstilboestrol. This suggested that ZER, a plant-derived compound, could be introduced as a new chemo-preventive agent for CIN in future.

Topics: Animals; Antineoplastic Agents; Apoptosis; Carcinogens; Diethylstilbestrol; Female; Immunohistochemistry; In Situ Nick-End Labeling; Mice; Mice, Inbred BALB C; Microscopy, Confocal; Mitochondria; Pregnancy; Prenatal Exposure Delayed Effects; Reverse Transcriptase Polymerase Chain Reaction; Sesquiterpenes; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Recent in vitro and in vivo studies have demonstrated that zerumbone (ZER) possesses anticancer properties. The main objective of this study was to examine the effectiveness of the combination of ZER and cisplatin (CIS) to treat cervical intraepithelial neoplasia (CIN) in vivo. Microculture tetrazolium assay and immunohistochemistry of proliferating cellular nuclear antigen were used to study the antitumor effect of ZER. Prenatally exposed female BALB/c mice were used as a model. The progenies with CIN were injected peritoneally with isotonic sodium chloride solution (positive control), CIS, ZER, and a combination of both compounds. All treated and untreated mice were humanely killed, and serum and cervix were obtained for interleukin 6 analysis and histopathologic studies using hematoxylin and eosin staining, respectively. Zerumbone has revealed an antitumor effect on human cervical cancer cells and downregulates immunoexpression of proliferating cellular nuclear antigen (P < 0.05). In vivo study indicates that ZER at 16 mg/kg and CIS at 10 mg/kg have a regressing effect on CIN. The combination of ZER and CIS also showed similar effectiveness in regressing CIN. Our results indicate that the combination of ZER and CIS has modulated the serum level of interleukin 6 when compared with that in mice treated with isotonic sodium chloride solution (P < 0.05). The effectiveness of combining ZER and CIS could be further explored as a new therapeutic intervention of early precancerous stages of carcinogenesis before the invasive stage begins.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Cell Proliferation; Cell Survival; Cisplatin; Diethylstilbestrol; Disease Models, Animal; Drug Screening Assays, Antitumor; Female; HeLa Cells; Humans; Male; Mice; Mice, Inbred BALB C; Sesquiterpenes; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms