crocin and Uterine-Cervical-Neoplasms

Cervical cancer is the most common malignant tumor in females worldwide. Human papillomavirus (HPV) infection is associated with the occurrence of cervical cancer. Thus, developing an effective and low-cost vaccine against HPV infection, especially in developing countries is an important issue. In this study, a novel HPV L1-E7 fusion multiepitope construct designed by immunoinformatics tools was expressed in bacterial system. HEK-293T cells-derived exosomes were generated and characterized to use as a carrier for crocin and curcumin compounds. The exosomes loaded with crocin and curcumin compounds as a chemotherapeutic agent (ExoCrocin and ExoCurcumin) were used along with the L1-E7 polypeptide for evaluation of immunological and anti-tumor effects in C57BL/6 mouse model. In vitro studies showed that ExoCrocin and ExoCurcumin were not cytotoxic at a certain dose, and they could enter tumor cells. In vivo studies indicated that combination of the L1-E7 polypeptide with ExoCrocin or ExoCurcumin could produce a significant level of immunity directed toward Th1 response and CTL activity. These regimens showed the protective and therapeutic effects against tumor cells (the percentage of tumor-free mice: ~100%). In addition, both ExoCrocin and ExoCurcumin represented similar immunological and anti-tumor effects. Generally, the use of exosomal crocin or curcumin forms along with the L1-E7 polypeptide could significantly induce T-cell immune responses and eradicate tumor cells.

Topics: Animals; Capsid Proteins; Carotenoids; Cell Line, Tumor; Curcumin; Drug Synergism; Exosomes; Female; HEK293 Cells; Humans; Mice; Mice, Inbred C57BL; Oncogene Proteins, Viral; Papillomavirus E7 Proteins; Papillomavirus Vaccines; Peptides; Recombinant Proteins; Treatment Outcome; Uterine Cervical Neoplasms

The present study was aimed to investigate anticancer activity of crocin against cervical carcinoma and bio-assessment and toxicological evaluation in male albino rats. Effect of crocin on cell viability (anticancer activity) was determined against cervical carcinoma cells. Chronic effect of crocin on body weight changes, serum enzymes, serum biochemical markers, lipid peroxidation, hematological markers and DNA damage in male albino rats were determined. Cell survival rate was reduced 98.4, 95.7, 87.2, 81.1 and 73.1% at 25, 50, 75, 100 and 125 mg/l of crocin respectively. Cell viability was reduced 97.1, 96.4, 85.5, 78.4 and 70.2% at 25, 50, 75, 100 and 125 mg/l of crocin respectively. Crocin reduced body weight significantly at 30 and 60th day. Alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), blood urea nitrogen (BUN), creatinine, bilirubin, albumin and total protein were decreased, while glucose, cholesterol, TG, and GSH were increased. Hemoglobin (Hb), white blood cells (WBC), lymphocytes, neutrophil and packed cell volume (PCV) were altered following crocin treatment. Necrosis, fibrosis, mononuclear infiltration, angiogenesis and DNA fragmentation were also noted. Taking all these data together, it is suggested that the crocin could be a potential antitumor agent against cervical carcinoma. However, the altered histological, biochemical and hematological markers may lead to an adverse effect on the cellular metabolism and physiological activity.

Topics: Administration, Oral; Alanine Transaminase; Alkaline Phosphatase; Animals; Antineoplastic Agents; Aspartate Aminotransferases; Biomarkers; Body Weight; Brain; Carotenoids; Cell Survival; DNA Fragmentation; Female; HeLa Cells; Humans; Liver; Male; Malondialdehyde; Rats; Rats, Wistar; Uterine Cervical Neoplasms

Cervical cancer is the fourth cause of cancer-related mortality among females worldwide. Although current therapies reduce disease symptoms, resistance of tumor cells to chemotherapy agents after a while is a serious problem. Therefore, utilization of novel adjuvant agents to increase efficiency of chemotherapy is essential. In the last two decades, botanicals with effective anticancer activities have been studied. Among them, the anticancer properties of crocin have been more attended. In this study, the molecular mechanism of crocin action was investigated in sensitive human cervical cancer cell line (OV2008) in comparison with the resistant one (C13). A 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay showed that crocin inhibits proliferation of sensitive cells (OV2008) at a time- and dose-dependent manner at 48 and 72h. Also, this inhibitory effect has been shown on resistant cells (C13) at 72h. Hoechst staining and flow cytometry assay also confirmed these results and revealed that antiproliferative effect of crocin might be due to the induction of apoptosis. Moreover, the genetic mechanism of crocin-induced apoptosis was accomplished by studying the relative expressions of P53, Bax, Bcl2 and miR-365, an upstream regulator of the last two ones. Real-time PCR analysis indicated that 1.5 and 3mg/ml crocin led to up-regulation of Bax and P53 and down-regulation of Bcl2 and miR-365 at all time intervals in both two cell lines. However, OV2008 cell line was more sensitive to crocin, and alternation of gene expretion was more obvious in this cell line. In this regard, the present study demonstrated the anti-proliferative and apoptotic activities of crocin against both sensitive and resistant cervical cancer cells that may benefit cervical cancer treatment as an adjuvant agent to decrease chemoresistance and increase the efficiency of therapy.

Topics: Antineoplastic Agents; Apoptosis; Carotenoids; Cell Culture Techniques; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Female; Humans; Uterine Cervical Neoplasms