One‐Pot Tandem [4 + 4] Annulation of 2‐Fluoro‐N‐arylbenzenesulfonamides with 2‐Methylidenetrimethylene Carbonates: Access to Functionalized Benzoxathiazocines

Cytoplasmic NANOG-Positive Stromal Cells Promote Human Cervical Cancer Progression

Cervical and breast cancer are some of the deadliest forms of cancer that may occur in women. According to the Indonesian Ministry of Health, cervical and breast cancer makes up 0.8% of all cancer cases in Indonesia. Conventional treatment methods such as chemotherapy have many disadvantages such as stomach problems, hair loss, darkening of the skin, and nausea. Therefore, a form of therapy that specifically targets cancer cells has been a hot topic in novel researches. The aim of this research explores the effect of propolis from Indonesian bee Tetragonula biroi on human cancer cell lines HeLa and MCF-7. Ethanolic extract of Propolis was obtained from raw propolis. 250 ppm of the samples was added to the cell lines. Subsequently, propolis’ activity in inhibiting cell growth was analyzed using the MTT Assay method. The inhibition percentage was then obtained. The propolis extracts were also analyzed by LC-MS/MS to identify anticancer components that exist, also with Spectrophotometer UV-Vis to identify the amount of flavonoids and polifenols present in the extract. The results show that rough propolis has the higher cytotoxic effect with inhibition percentages of 92.42% for MCF-7 and 86.81% for HeLa cells, while smooth propolis inhibits MCF-7 growth by 87.60% and HeLa by 77.27%. It may be concluded in this study that while propolis has potential as an anticancer agent, future research is still very much needed.

BackgroundSeveral reports have revealed that cancer stem cells (CSCs) exist in many types of solid tumors. Some studies have demonstrated that side population (SP) cells isolated from diverse cancer lines harbor cancer stem-like properties, but there are few reports examining the characteristic of SP cells in human cervical cancer. The aim of this study is 1) to find out a feasible way to detect the tumor stem-like cells in cervical cancer, and 2) to analyze the properties of the SP cells being sorted.MethodsIsolated SP and non-SP cells from human cervical cancer cell line Hela by Hoechst 33342 dying method and flow cytometry analysis. Observing morphology of SP and non-SP cells. The expression of various biomarkers putatively related to cancer stem cells were investigated by immucytochemistry of SP and non-SP cells. We also analyzed cell cycle and cell apoptosis for sorted cells. The oncogenicity of the SP and non-SP cells were analyzed by tumor formation in nonobesediabeti- c/severe combined immune- deficient (NOD/SCID) mice. The drug-resistant and radiation-resistant index between SP, non-SP and Hela cells was estimated by MTS assay.ResultsThe fraction of SP cells in Hela was approximately 1.07 ± 0.32%. SP cells were smaller and rounder in shape than non-SP cells, and mostly showed colony-like growth. Immunocytochemistry showed that stem cell makers (Oct3/4, CD133, BCRP) were highly expressed in SP cells. Moreover, the number of apoptotic cells among non-SP cells (17.6 ± 3.7%) was significantly higher compared with that among SP cells (4.4 ± 1.2%). The HE staining of in vivo grown tumors result from SP cells showed more poor differentiation, though no significant differences were shown between SP and non-SP cells in NOD/SCID mice tumorigenicity. Furthermore, SP cells demonstrated a higher degree of drug resistance against trichostatin A (TSA) compared with that of non-SP and Hela cells. SP cells were also found to be more resistant against radiotherapy.ConclusionsSP cells possess some characteristics of CSCs, namely high proliferation ability, chemoresistance and radioresistance, which may be helpful to elucidate novel targets for effective clinical treatments of cervical cancer in the future.

Cervical cancer is one of the deadliest cancers in women with a death toll of 230,000 worldwide each year, nearly 80% in developing countries. Radiotherapy (RT) is a major treatment modality for advanced cervical cancer but the local relapse rate is 30-44% in patients treated with RT alone and 19-25% in patients treated with concurrent chemoradiotherapy. Previous studies have shown that the transcription factor NF-kappaB is constitutively expressed in human cervical squamous cell carcinomas. NF-kappaB activation also contributes to the resistance of cervical cancer cells to apoptosis induced by chemotherapeutic agents and radiation. Therefore, inhibition of NF-kappaB in tumor cells may render them more sensitive to chemo/radiation therapies. The objective of this study is to investigate the potential of radiosensitization of NF-kappaB inhibition by Velcade in human cervical cancer cell lines. We used the human cervical cancer cell lines HeLa and SiHa. Both are highly radioresistant and chemoresistant as compared to other cervical cancer cell lines. These cells had been treated with Velcade before they were irradiated with different doses of ionizing radiation. MTT metabolic assays and clonogenic cell survival assays were performed to evaluate the effects of Velcade on radiation resistance. Inhibition of NF-kappaB by Velcade alone decreased metabolic potential (MTT) and clonogenic survival in SiHa, but not in HeLa cells. Furthermore, pre-treatment of SiHa, but not HeLa cells with Velcade enhanced radiation sensitivity. Inhibition of NF-kappaB by the proteasome inhibitor Velcade enhances radiosensitivity of certain human cervical carcinoma cancer cells in vitro. These results raise the possibility that inhibition of NF-kappaB will result in radiosensitization only in those tumor cells which are more dependent on NF-kappaB for their metabolism and survival, however, the radiosensitivity of "NF-kappaB independent" cells are not likely influenced by it.

Cervical cancer is the fourth most prevalent cancer among women worldwide and remains a significant contributor to cancer-related mortality, particularly in low- and middle-income countries, largely due to treatment resistance and disease recurrence. Growing evidence indicates that cancer stem cells (CSCs) play a pivotal role in tumor initiation, progression, and therapeutic resistance. Consequently, the development of novel therapeutic strategies capable of eliminating cancer cells while simultaneously targeting CSC-associated pathways holds substantial clinical promise. This study aimed to evaluate the combined effects of zinc phthalocyanine-mediated photodynamic therapy (ZnPc-PDT) combined with oxaliplatin on cervical cancer cells. The focus was on assessing cell viability, apoptosis, colony formation, migration, stemness characteristics, and the expression of key genes involved in CSC regulation. Human cervical cancer cell lines (HeLa and Caski) were treated with ZnPc-PDT, oxaliplatin, or their combination. Cytotoxicity was measured using MTT assays, while apoptosis was evaluated by Annexin V/PI flow cytometry and expression profiling of apoptosis-related genes (CASPASE3, CASPASE8, CASPASE9, BCL2). Colony-forming assays were used to assess stemness potential, and wound-healing assays evaluated cell migration. Quantitative real-time PCR (qRT-PCR) was performed to examine the expression of stemness-related markers (SOX2, OCT4, CD133, CD44) and metastasis-associated genes (MMP2, MMP9, ROCK1). Additionally, in silico pathway analysis using TCGA-CESC, STRING, and Enrichr datasets identified oxaliplatin-targeted genes involved in CSC regulation and validated the experimental observations. Human cervical cancer cell lines (HeLa and Caski) were treated with ZnPc-PDT, oxaliplatin, or their combination. Cytotoxicity was measured using MTT assays, while apoptosis was evaluated by Annexin V/PI flow cytometry and expression profiling of apoptosis-related genes (CASPASE3, CASPASE8, CASPASE9, BCL2). Colony-forming assays were used to assess stemness potential, and wound-healing assays evaluated cell migration. Quantitative real-time PCR (qRT-PCR) was performed to examine the expression of stemness-related markers (SOX2, OCT4, CD133, CD44) and metastasis-associated genes (MMP2, MMP9, ROCK1). Additionally, in silico pathway analysis using TCGA-CESC, STRING, and Enrichr datasets identified oxaliplatin-targeted genes involved in CSC regulation and validated the experimental observations. The combination of ZnPc-PDT and oxaliplatin exhibits potent anti-cancer effects by inducing apoptosis, suppressing migration, reducing stemness, and modulating key cancer-related pathways. By integrating molecular experiments with in silico analysis, this study provides mechanistic insights into how PDT enhances oxaliplatin efficacy. These findings suggest that ZnPc-PDT combined with oxaliplatin may represent a promising therapeutic strategy to overcome drug resistance and reduce recurrence in cervical cancer treatment.

Novel diastereoselective trans 2, 3-dihydrobenzofuran derivatives: Tandem synthesis, crystal structure, antioxidant and anticancer activity

A three-component cascade method has been developed for the direct synthesis of polysubstituted pyridines. This strategy provides a very convenient route to pyridines using a variety of β-bromo-α,β-unsaturated aldehydes, 1,3-diketones, and ammonium acetate without any additional catalyst or metal salt under mild conditions. A variety of β-ketoesters and 4-hydroxycoumarin were also used instead of 1,3-diketones for the diverse synthesis of polycyclic pyridines. One of the synthesized pyridines has been unambiguously established by a single crystal XRD study. All of the synthesized pyridine derivatives were evaluated for their antiproliferative properties in vitro against the human cancer cell lines HeLa, Me180, and ZR751. Compounds 4{4,1} and 4{2,4} showed significant cytotoxicity in the human breast cancer cell line ZR751 and cervical cancer cell line Me180, respectively, and a few other compounds were found to have moderate activities.

In this study, RNA-sequencing was used to investigate the differentially expressed miRNAs between cervical cancer tissues and matched adjacent non-tumor tissues. Five miRNAs were sharply downregulated in the cancer tissue, including miR-199a, miR-22, miR-615, miR-3681-3p (miR-3681), and miR-1193. Among them, miR-3681 was uncharacterized. The results from qPCR analysis showed that miR-3681 expression was decreased in patients with cervical cancer compared with the control, and decreased in the human cervical cancer cell lines SiHa, HeLa, C4-1, C-33A and Caski, compared with the normal human cervical epithelial cell line HCerEpic. Then, different concentrations of miR-3681 mimic and miR-3681 inhibitor were respectively transfected into the human cervical cancer cell line C-33A, and the expression of miR-3681, cell proliferation, cell apoptosis and cell migration were measured after 48 h. The results showed that the miR-3681 mimic increased the miR-3681 level, suppressed cell proliferation and migration, and induced cell apoptosis in a dose-dependent manner. In contrast, the miR-3681 inhibitor decreased the miR-3681 level, promoted cell proliferation and migration, and inhibited cell apoptosis in a dose-dependent manner. Moreover, bioinformatics analysis showed that there was a miR-3681 binding site in the mRNA 3′UTR of HGFR, which was robustly upregulated in cervical cancer cell lines compared with HCerEpic cells. In addition, luciferase activity analysis demonstrated that miR-3681 could directly target HGFR, which promoted the proliferation and migration of C-33A cells via activation of the PI3K/Akt pathway in a dose-dependent manner. Furthermore, our results showed that knockdown of HGFR could antagonize the promotion of anti-miR-3681 on the activation of the PI3K/Akt pathway and cell proliferation and migration. In conclusion, MiR-3681 was identified as a negative regulator in the proliferation and migration of cervical cancer cells. This function is associated with the posttranscriptional suppression of HGFR and the deactivation of the PI3K/Akt pathway.

Synthesis and biological evaluation of spiro[cyclopropane-1,3'-indolin]-2'-ones as potential anticancer agents.

To explore the mechanism of miR-202-5p targeting the expression of PIK3CA and mediating the activation of PI3K/Akt/mTOR signaling pathway on the proliferation, invasion, and epithelial-mesenchymal transition (EMT) of cervical cancer. The objects of study were 105 cases of cervical cancer and their corresponding normal tissues. qRT-PCR was used to detect the expression of miR-202-5p and PIK3CA in adjacent normal tissue and cervical cancer tissue. Dual luciferase reporter assay was used to verify the targeting relationship between miR-202-5p and PIK3CA gene. Human cervical cancer cell lines HPV-16E6, SiHa, HeLa, and CaSki were purchased for our cell experiments. The expression levels of PIK3CA in the cells were detected by qRT-PCR. The cell line with higher expression levels was selected to complete the follow-up experiment. The cultured cells were transfected and divided into the miR-202-5p mimic NC group, miR-202-5p mimic group, miR-202-5p inhibitor NC group, miR-202-5p inhibitor group, siRNA-PIK3CA NC group, siRNA-PIK3CA group, miR-202-5p inhibitor NC + siRNA-PIK3CA NC group, miR-202-5p inhibitor + siRNA-PIK3CA NC group, and miR-202-5p inhibitor + siRNA-PIK3CA group. QRT-PCR was used to detect the expression of miR-202-5p. Western blot and qRT-PCR were applied to detect the mRNA and protein expression levels of related pathway proteins (PIK3CA, PI3K, PTEN, p-Akt1, and p-mTOR) and epithelial-mesenchymal transition-related factors (N-cadherin, E-cadherin, and vimentin). Cell proliferation was detected by plate colony formation assay. Transwell assay was used to detect the invasion ability of each group. When compared with the adjacent tissues, PIK3CA mRNA expression level was significantly increased and miR-202-5p expression level was significantly decreased in cervical cancer tissues (all P < 0.05). PIK3CA was a target gene of miR-202-5p. The mRNA expression level of PIK3CA in SiHa cervical cancer cells was significantly higher than that in CaSki, HeLa, and HPV-16E6 cells (all P < 0.05), and SiHa cervical cancer cells were selected to complete the follow-up experiments. When compared with the corresponding NC group, the expression of miR-202-5p in miR-202-5p mimic group was increased. In addition, the mRNA and protein expression levels of E-cadherin and PTEN in miR-202-5p mimic and siRNA-PIK3CA groups were increased, and the protein expression of p-Akt1 and p-mTOR was decreased, and also, the mRNA and protein expression levels of PIK3CA, PI3K, N-cadherin, and vimentin were decreased (all P < 0.05); in miR-202-5p inhibitor group, the expression levels of miR-202-5p, E-cadherin, and PTEN decreased, the protein expression of p-Akt1 and p-mTOR increased, and the mRNA and protein expression of PIK3CA, PI3K, N-cadherin, and vimentin increased in miR-202-5p inhibitor group (all P < 0.05); in miR-202-5p inhibitor + siRNA-PIK3CA group, the expression of miR-202-5p decreased (P < 0.05), but the mRNA and protein expression of PIK3CA, PI3K, p-Akt1, p-mTOR, N-cadherin, E-cadherin, and vimentin had no significant changes (all P > 0.05). When compared with the corresponding NC group, the number of cell clones in miR-202-5p mimic group and siRNA-PIK3CA group was decreased, and the invasion ability of miR-202-5p inhibitor group was increased, and the invasion ability was enhanced (all P < 0.05); miR-202-5p inhibitor + siRNA-PIK3CA group showed no significant change in the number of cell clones and the rate of invasion (P > 0.05). In conclusion, the overexpression of miR-202-5p can suppress PIK3CA gene expression and the activation of PI3K/Akt/mTOR signaling pathway to suppress the proliferation, invasion, and EMT of cervical cancer.

Synaptonemal complex protein 3 (SCP3), a member of Cor1 family, is up-regulated in various cancer cells; however, its oncogenic potential and clinical significance has not yet been characterized. In the present study, we investigated the oncogenic role of SCP3 and its relationship with phosphorylated AKT (pAKT) in cervical neoplasias. The functional role of SCP3 expression was investigated by overexpression or knockdown of SCP3 in murine cell line NIH3T3 and human cervical cancer cell lines CUMC6, SiHa, CaSki, and HeLa both in vitro and in vivo. Furthermore, we examined SCP3 expression in tumor specimens from 181 cervical cancer and 400 cervical intraepithelial neoplasia (CIN) patients by immunohistochemistry and analyzed the correlation between SCP3 expression and clinicopathologic factors or survival. Overexpression of SCP3 promoted AKT-mediated tumorigenesis both in vitro and in vivo. Functional studies using NIH3T3 cells demonstrated that the C-terminal region of human SCP3 is important for AKT activation and its oncogenic potential. High expression of SCP3 was significantly associated with tumor stage (P = 0.002) and tumor grade (P<0.001), while SCP3 expression was positively associated with pAKT protein level in cervical neoplasias. Survival times for patients with cervical cancer overexpressing both SCP3 and pAKT (median, 134.0 months, n = 68) were significantly shorter than for patients with low expression of either SCP3 or pAKT (161.5 months, n = 108) as determined by multivariate analysis (P = 0.020). Our findings suggest that SCP3 plays an important role in the progression of cervical cancer through the AKT signaling pathway, supporting the possibility that SCP3 may be a promising novel cancer target for cervical cancer therapy.

Cervical cancer is a major challenge facing women's health worldwide. In China, its incidence and mortality rate rank first among female reproductive tract malignancies. SNHG15 has been found and proven to have important functions in many cancers. Related studies have shown that miR-200a-3p plays an important role in tumor development. However, there is little research on how the two are involved in the progression of cervical cancer cells and their potential regulatory systems. qRT-PCR was used to detect the expression of LncRNA SNHG15 in human cervical immortalized squamous cells (Ect1/E6E7) and human cervical cancer cell lines (SiHa, HeLa, Caski, C-33A). Cervical cancer HeLa and SiHa cells were used as the research objects and were divided into NC, GV493-negative group, SNHG15-shRNA group, empty vector group, and SNHG15-OERNA group. The expression levels of LncRNA SNHG15 and miR-200a-3p in cells were detected by qRT-PCR. CCK8 was used to detect cell proliferation, cell migration and invasion ability, and dual luciferase activity was used to analyze the targeted binding of LncRNA SNHG15 and miR-200a-3p. Compared with human cervical immortalized squamous cells (Ect1/E6E7), the expression level of LncRNA SNHG15 in human cervical cancer cell lines SiHa, HeLa and C-33A was increased(P<0.05). Cervical cancer cells HeLa and SiHa with more significant differences were selected for subsequent studies. Overexpression of LncRNA SNHG15 or low expression of miR-200a-3p promoted cell proliferation, migration and invasion (P < 0.05). High expression of SNHG15 progression cervical cancer cell proliferation, migration, and invasion by targeting the miR-200a-3p gene. Trial registration: DRKS, ISRCTNDRKS00035987. Registration on 17 February 2025-Retrospectively registered, https://drks.de/register/de/trial/DRKS00035987.

Background: Tagetes species have been extensively employed in folk medicine and have been demonstrated to exert various biological activities due to its phytochemical constituents, including those related to cancer. Objectives: The aim of this study was to identify, in Tagetes lucida organic leaf extracts, the bioactive compounds with antiproliferative and antimigratory activities on the human cervical cancer (CCa) cell lines human cervical cancer cell line (HPV-16) (SiHa) and human cervical cancer cell line (HPV-18) (HeLa), and on the non-cancer cell line human immortalized keratinocyte cell line (HaCaT). Materials and Methods: The phytochemical profile of all Tagetes leaves extracts was determined by Gas chromatography mass spectroscopy analysis and tested on SiHa, HeLa, and HaCaT cell lines using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and wound-healing assays. Results: The Gas chromatography coupled with mass analysis analysis revealed two main constituents identified in all extracts, the coumarins herniarin (17.152%–56.904% content) and scoparone (2.778%–34.817% content), the rest of identified constituents were terpenes where the geranyl acetate was the major constituent on hexane extract. On the anti-proliferative potential of T. lucida extracts, our present investigation revealed that all extracts decreased HeLa cell viability (half-maximal inhibitory concentration value of <190.26 mg/mL at 24 h and <220.41 mg/mL at 48 h) dose dependently; the cell viability of SiHa and HaCaT was not affected. Furthermore, the migration capacity of SiHa and HeLa cells decreased after 24 and 48 h with acetonic and methanolic extracts treatment, while on HeLa cells a more evident effect was observed with a <80% decrease in wound closure. Conclusion: Our findings revealed the coumarins presented in T. lucida as potential candidates of inhibiting cell proliferation and migration preferably toward CCa (HeLa cell line).

Red berries are important sources of bioactive compounds and they are known to provide unique health benefits. Lately, it has been proved that anthocyanins have health benefits against degenerative diseases such as cardiovascular disease, cancer or diabetes. Accordingly, the aim of this study was to characterize the anthocyanin content of anthocyanins pure extracts (APEs) obtained from raspberries (Rubus sp.) and mulberries (Morus sp.) and to evaluate their antiproliferative effect in vitro. Upon chromatographic analysis, three anthocyanins were identified in purified extracts of mulberries (M-APEs), with cyanidin-3-O-glucoside being more abundant. On the other hand, purified extracts of raspberries (R-APEs) contained 2 anthocyanins, both identified as cyanidin-derivatives. The in vitro test demonstrated that APEs decreased the proliferation on both HeLa and A2780 human cancer cell lines in a dose dependent manner, demonstrating that these two different berries are both rich sources of anthocyanins and are able to exert antiproliferative proprieties toward cervical and ovarian cancer.

Convenient and efficient synthesis of novel N-(4-acetyl-4,5-dihydro-5-(7,8,9-substituted-tetrazolo[1,5-a]-quinolin-4-yl)-1,3,4-thiadiazol-2-yl)acetamides 4a–j and their in vitro anticancer activity against two cell lines viz., human breast cancer cell line MCF7 and human cervix cancer cell line HeLa were carried out. GI50, LC50, TGI values were evaluated. Two of the compounds 4e and 4i with halogen substituent at 7th position of the target molecules have shown potent activity against human cervix cancer cell line HeLa. DNA cleavage studies revealed that most of these compounds show partial cleavage and few of them show complete cleavage of DNA.