Deoxyshikonin triggers apoptosis in cervical cancer cells through p38 MAPK-mediated caspase activation.

Deoxyshikonin (DSK) is a biological component derived from Lithospermum erythrorhizon. Although DSK possesses potential anticancer activities, whether DSK exerts anticancer effects on cervical cancer cells is incompletely explored. This study was aimed to investigate the anticancer activity of DSK against cervical cancer cells and its molecular mechanisms. Cell viability was evaluated by MTT assay. Level of phosphorylation and protein was determined using Western blot. Involvement of signaling kinases was assessed by specific inhibitors. Our results revealed that DSK reduced viability of human cervical cell in a dose-dependent fashion. Meanwhile, DSK significantly elicited apoptosis of HeLa and SiHa cells. Apoptosis microarray was used to elucidate the involved pathways, and the results showed that DSK dose-dependently diminished cellular inhibitor of apoptosis protein 1 (cIAP1), cIAP2, and XIAP, and induced cleavage of poly(ADP-ribose) polymerase (PARP) and caspase-8/9/3. Furthermore, we observed that DSK significantly triggered activation of ERK, JNK, and p38 MAPK (p38), and only inhibition of p38 diminished the DSK-mediated pro-caspases cleavage. Taken together, our results demonstrate that DSK has anti-cervical cancer effects via the apoptotic cascade elicited by downregulation of IAPs and p38-mediated caspase activation. This suggests that DSK could act as an adjuvant to facilitate cervical cancer management.

Impact of LINC00673 genetic variants on uterine cervical cancer clinicopathologic characteristics.

To date, no study delineates the relationships among the genetic variants of long intergenic noncoding RNA 673 (LINC00673) and uterine cervical carcinogenesis as well as clinicopathological parameters and 5 years survival of cervical cancer patients in Taiwan. Therefore, the involvement of LINC00673 polymorphisms in cervical cancer was investigated. Genotypic frequencies of three LINC00673 polymorphisms rs6501551, rs9914618 and rs11655237 were determined in 199 patients including 115 patients with invasive cancer, 84 with precancerous lesions, and 274 control females using real-time polymerase chain reaction. It revealed that LINC00673 polymorphisms were not found significantly related to development of cervical cancer. Cervical cancer patients with genotypes AG/GG in LINC00673 rs6501551 had more risk to have tumor diameter larger than 4 cm as compared to those with genotype AA (p=0.043). Cervical cancer patients with genotype GG in rs6501551 had worse 5 years survival as compared to those with genotypes AA/AG in multivariate analysis (hazard ratio: 4.70; p=0.097). However, only two patients exhibiting GG were noted, and one had mortality, another had no mortality. In conclusion, larger sample size needs to verify the associations of LINC00673 genetic variants with clinicopathological parameters and patient survival of cervical cancer for Taiwanese females.

The most likely but largely ignored triggering factor for breast (or all) cancer invasion.

Breast cancer development and progression are believed to be a sequential process, from normal to hyperplastic, to in situ, and to invasive and metastatic stages. Given that over 90% of cancer deaths are caused by invasive and metastatic lesions, countless factors and multiple theories have been proposed as the triggering factor for the cascade of actions of cancer invasion. However, those factors and theories are largely based on the studies of cell lines or animal models. In addition, corresponding interventions based on these factors and theories have failed to reduce the incidence rate of invasive and metastatic lesions, suggesting that previous efforts may have failed to arm at the right target. Considering these facts and observations, we are proposing "A focal aberrant degeneration in the myoepithelial cell layer (MECL) as the most likely triggering factor for breast cancer invasion". Our hypothesis is based on our recent studies of breast and multiple other cancers. Our commentary provides the rationale, morphologic, immunohistochemical, and molecular data to support our hypotheses. As all epithelium-derived cancers share a very similar architecture, our hypothesis is likely to be applicable to invasion of all cancer types. We believe that human tissue-derived data may provide a more realistic roadmap to guide the clinic practice.

Laser-assisted failure recovery for dielectric elastomer actuators in aerial robots.

Insects maintain remarkable agility after incurring severe injuries or wounds. Although robots driven by rigid actuators have demonstrated agile locomotion and manipulation, most of them lack animal-like robustness against unexpected damage. Dielectric elastomer actuators (DEAs) are a class of muscle-like soft transducers that have enabled nimble aerial, terrestrial, and aquatic robotic locomotion comparable to that of rigid actuators. However, unlike muscles, DEAs suffer local dielectric breakdowns that often cause global device failure. These local defects severely limit DEA performance, lifetime, and size scalability. We developed DEAs that can endure more than 100 punctures while maintaining high bandwidth (>400 hertz) and power density (>700 watt per kilogram)-sufficient for supporting energetically expensive locomotion such as flight. We fabricated electroluminescent DEAs for visualizing electrode connectivity under actuator damage. When the DEA suffered severe dielectric breakdowns that caused device failure, we demonstrated a laser-assisted repair method for isolating the critical defects and recovering performance. These results culminate in an aerial robot that can endure critical actuator and wing damage while maintaining similar accuracy in hovering flight. Our work highlights that soft robotic systems can embody animal-like agility and resilience-a critical biomimetic capability for future robots to interact with challenging environments.

CLEFMA induces intrinsic and extrinsic apoptotic pathways through ERK1/2 and p38 signalling in uterine cervical cancer cells.

Although concurrent chemoradiotherapy is the cornerstone of treatment for locally advanced or recurrent uterine cervical cancer, treatment fails at a high rate. Therefore, the development of novel targeting agents is critical. This study investigated the action of CLEFMA, a potent, synthetic curcumin derivative, on cervical cancer cells and its mechanism of action. We found that CLEFMA negatively regulated the viability of cervical cancer cells, involving induction of cell apoptosis. Cleaved caspase-3, cleaved poly(adenosine diphosphate-ribose) polymerase, cleaved caspase-8, and cleaved caspase-9 expression were increased by treatment with CLEFMA. After U0126 (ERK1/2 inhibitor) and SB203580 (p38 inhibitor) were applied as cotreatment with CLEFMA, the expression of cleaved caspase-8, -9, and -3 was reduced significantly. In conclusion, CLEFMA activates both extrinsic and intrinsic apoptotic pathways through ERK1/2 and p38 signal transduction in cervical cancer cells.

Synergistic Combination of Luteolin and Asiatic Acid on Cervical Cancer In Vitro and In Vivo.

Cervical cancer is an important issue globally because it is the second most common gynecological malignant tumor and conventional treatment effects have been shown to be limited. Lut and AsA are plant-derived natural flavonoid and triterpenoid products that have exhibited anticancer activities and can modulate various signaling pathways. Thus, the aim of the present study was to evaluate whether Lut combined with AsA could enhance the anticancer effect to inhibit cervical cancer cell proliferation and examine the underlying molecular mechanisms in vitro and in vivo. The results of a CCK-8 assay showed that Lut combined with AsA more effectively inhibited the proliferation of CaSki and HeLa cells than Lut or AsA treatment alone. Lut combined with AsA caused apoptosis induction and sub-G1-phase arrest in CaSki and HeLa cells, as confirmed by flow cytometry, mitoROS analysis, antioxidant activity measurement and western blot assay. In addition, Lut combined with AsA significantly inhibited the cell migration ability of CaSki and HeLa cells in a wound-healing assay. Furthermore, Lut combined with AsA induced apoptosis and inhibited migration through downregulated PI3K/AKT (PI3K, AKT and p70S6K), JNK/p38 MAPK and FAK (integrin ß1, paxillin and FAK) signaling and upregulated ERK signaling. In an in vivo study, Lut combined with AsA markedly inhibited cervical cancer cell-derived xenograft tumor growth. Collectively, the present study showed that Lut combined with AsA may be used as an anticancer agent to improve the prognosis of cervical cancer. Indeed, with additional research to develop standardized dosages, Lut and AsA combination therapy could also be applied in clinical medicine.

EF-24 inhibits TPA-induced cellular migration and MMP-9 expression through the p38 signaling pathway in cervical cancer cells.

Diphenyl difluoroketone (EF-24), a synthetic curcumin analog, has enhanced bioavailability over curcumin. EF-24 acts more powerful bioactivity for anti-inflammatory and anti-cancer activity. However, the effects and mechanism of EF-24 on cervical cancer has not been fully investigated. Herein, this study evaluated the effects of EF-24 on TPA-induced cellular migration of cervical cancer. The results showed that EF-24 substantially reduced the cellular migration and cellular invasion of the HeLa and SiHa cells. Moreover, gelatin zymography, western blotting analyses and real-time PCR revealed that EF-24 suppressed Matrix metalloproteinase-9 (MMP-9) activity, protein expression and mRNA levels. Mechanistically, EF-24 inhibited the phosphorylation of the p38 signaling pathway. In conclusion, EF-24 inhibited TPA-induced cellular migration and cellular invasion of cervical cancer cell lines through modulating MMP-9 expression via downregulating signaling p38 pathway and EF-24 may have potential to serve as a chemopreventive agent of cervical cancer.

Dihydromyricetin Inhibited Migration and Invasion by Reducing S100A4 Expression through ERK1/2/ß-Catenin Pathway in Human Cervical Cancer Cell Lines.

Cervical cancer has a poor prognosis and is the fourth most common cancer among women. Dihydromyricetin (DHM), a flavonoid compound, exhibits several pharmacological activities, including anticancer effects; however, the effects of DHM on cervical cancer have received insufficient research attention. This study examined the antitumor activity and underlying mechanisms of DHM on human cervical cancer. Our results indicated that DHM inhibits migration and invasion in HeLa and SiHa cell lines. Mechanistically, RNA sequencing analysis revealed that DHM suppressed S100A4 mRNA expression in HeLa cells. Moreover, DHM inhibited the protein expressions of ß-catenin and GSK3ß through the regulated extracellular-signal-regulated kinase (ERK)1/2 signaling pathway. By using the ERK1/2 activator, T-BHQ, reverted ß-catenin and S100A4 protein expression and cell migration, which were reduced in response to DHM. In conclusion, our study indicated that DHM inhibited cell migration by reducing the S100A4 expression through the ERK1/2/ß-catenin pathway in human cervical cancer cell lines.

The most effective but largely ignored target for prostate cancer early detection and intervention.

Over the past two decades, the global efforts for the early detection and intervention of prostate cancer seem to have made significant progresses in the basic researches, but the clinic outcomes have been disappointing: (1) prostate cancer is still the most common non-cutaneous cancer in Europe in men, (2) the age-standardized prostate cancer rate has increased in nearly all Asian and African countries, (3) the proportion of advanced cancers at the diagnosis has increased to 8.2% from 3.9% in the USA, (4) the worldwide use of PSA testing and digital rectal examination have failed to reduce the prostate cancer mortality, and (5) there is still no effective preventive method to significantly reduce the development, invasion, and metastasis of prostate cancer… Together, these facts strongly suggest that the global efforts during the past appear to be not in a correlated target with markedly inconsistent basic research and clinic outcomes. The most likely cause for the inconsistence appears due to the fact that basic scientific studies are traditionally conducted on the cell lines and animal models, where it is impossible to completely reflect or replicate the in vivo status. Thus, we would like to propose the human prostate basal cell layer (PBCL) as "the most effective target for the early detection and intervention of prostate cancer". Our proposal is based on the morphologic, immunohistochemical and molecular evidence from our recent studies of normal and cancerous human prostate tissues with detailed clinic follow-up data. We believe that the human tissue-derived basic research data may provide a more realistic roadmap to guide the clinic practice and to avoid the potential misleading from in vitro and animal studies.

Effect of tissue inhibitor of metalloproteinases-3 genetics polymorphism on clinicopathological characteristics of uterine cervical cancer patients in Taiwan.

Single nucleotide polymorphisms (SNPs) of tissue inhibitor of metalloproteinases-3 (TIMP-3) have been revealed to be related to various cancers. To date, no study explores the relationships between TIMP-3 polymorphisms and uterine cervical cancer. The purposes of this research were to investigate the associations among genetic variants of TIMP-3 and development and clinicopathological factors of uterine cervical cancer, and patient 5 years survival in Taiwanese women. The study included 123 patients with invasive cancer and 97 with precancerous lesions of uterine cervix, and 300 control women. TIMP-3 polymorphisms rs9619311, rs9862 and rs11547635 were checked and their genotypic distributions were determined by real-time polymerase chain reaction. It showed that women with genotypes CT/TT in rs9862 were found to display a higher risk of developing cervical cancer with moderate and poor cell differentiation. Moreover, it revealed that cervical cancer patients carrying genotypes CC in rs9619311 exhibited a poorer 5 years survival, as compared to those with TT/TC in Taiwanese women, using univariate analysis. In addition, pelvic lymph node metastasis was determined to independently predict 5 years survival in cervical cancer patients using multivariate analysis. Conclusively, TIMP-3 SNPs polymorphisms rs9619311 are related to cervical patient survival in Taiwanese women.

Impact of Genetic Variants of Long Noncoding RNA Metastasis-Associated Lung Adenocarcinoma Transcript 1 on Uterine Cervical Cancer.

Genetic variants of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1) have been reported to be associated with several cancers. Until now, no study reveals the associations between lncRNA MALAT1 polymorphisms and cervical cancer (CC). The objectives of this study were to explore the correlations among MALAT1 polymorphisms and occurrence and clinicopathological parameters of CC, as well as patient 5 years survival in Taiwanese women. The study recruited 116 patients with cervical invasive cancer and 89 patients with cervical precancerous lesions, as well as 268 non-cancer control women. LncRNA MALAT1 polymorphisms rs3200401, rs619586 and rs1194338 were selected and their genotypic frequencies were defined by real-time polymerase chain reaction. Our results revealed that there are no relationships between lncRNA MALAT1 genetic variants and occurrence of CC. The independent factor among lncRNA MALAT1 genetic variants and clinicopathological parameters were positive pelvic lymph node metastasis (p=0.001, HR: 10.94, 95% CI: 2.65-45.23). In conclusions, lncRNA MALAT1 genetic variants are not related to occurrence and clinicopathological characteristics of CC and patient 5 years survival in Taiwanese women. Pelvic lymph node metastasis could independently predict the patient 5 years survival among various MALAT1 polymorphisms and clinicopathological factors in CC.

Experimental evidence of auxeticity in ion implanted single crystal calcite.

We report initial experimental evidence of auxeticity in calcite by ion implanting (1010) oriented single crystalline calcite with Ar+ at room temperature using an ion energy of 400 keV and a dose of 1 × 1014 cm-2. Lattice compression normal to the substrate surface was observed, which is an atypical result for ion implanted materials. The auxetic behavior is consistent with predictions that indicate auxeticity had been predicted along two crystallographic directions including [1010]. Materials with a positive Poisson's ratio experience lattice expansion normal to the substrate surface when ion implanted, whereas lattice contraction normal to the surface is evidence of auxetic behavior. Triple-axis X-ray diffraction measurements confirmed the auxetic strain state of the implanted calcite substrates. Reciprocal space maps for the symmetric 3030 and asymmetric 1450 reflections revealed that the implanted region was fully strained (pseudomorphic) to the bulk of the substrate, as is typical with implanted single crystals. A symmetric (3030) ω:2θ line scan was used with X-ray dynamical diffraction simulations to model the strain profile and extract the variation of compressive strain as a function of depth normal to the substrate surface. SRIM calculations were performed to obtain a displacement-per-atom profile and implanted Ar+ concentration profile. It was found that the strain profile matches the displacement-per-atom profile. This study demonstrated the use of ion implantation and X-ray diffraction methods to probe mechanical properties of materials and to test predictions such as the auxeticity.

Anticancer Effects and Molecular Mechanisms of Apigenin in Cervical Cancer Cells.

Cervical cancer is the fourth most frequent malignancy in women. Apigenin is a natural plant-derived flavonoid present in common fruit, vegetables, and herbs, and has been found to possess antioxidant and anti-inflammatory properties as a health-promoting agent. It also exhibits important anticancer effects in various cancers, but its effects are not widely accepted by clinical practitioners. The present study investigated the anticancer effects and molecular mechanisms of apigenin in cervical cancer in vitro and in vivo. HeLa and C33A cells were treated with different concentrations of apigenin. The effects of apigenin on cell viability, cell cycle distribution, migration potential, phosphorylation of PI3K/AKT, the integrin ß1-FAK signaling pathway, and epithelial-to-mesenchymal transition (EMT)-related protein levels were investigated. Mechanisms identified from the in vitro study were further validated in a cervical tumor xenograft mouse model. Apigenin effectively inhibited the growth of cervical cancer cells and cervical tumors in xenograft mice. Furthermore, the apigenin down-regulated FAK signaling (FAK, paxillin, and integrin ß1) and PI3K/AKT signaling (PI3K, AKT, and mTOR), inactivated or activated various signaling targets, such as Bcl-2, Bax, p21cip1, CDK1, CDC25c, cyclin B1, fibronectin, N-cadherin, vimentin, laminin, and E-cadherin, promoted mitochondrial-mediated apoptosis, induced G2/M-phase cell cycle arrest, and reduced EMT to inhibit HeLa and C33A cancer cell migration, producing anticancer effects in cervical cancer. Thus, apigenin may act as a chemotherapeutic agent for cervical cancer treatment.

Arctiin Inhibits Cervical Cancer Cell Migration and Invasion through Suppression of S100A4 Expression via PI3K/Akt Pathway.

Arctiin, a lignan glycoside, is isolated from Arctium lappa L. The anticancer effects of arctiin have been demonstrated in several studies. However, no research has been conducted on the anti-migration effect of arctiin in cervical cancer cells. The present study examined the effects of arctiin on cervical cancer cells and investigated the possible molecular mechanism. We demonstrated that arctiin exhibited low cytotoxicity and significantly inhibited cell migration and invasion in human cervical cancer cells. The S100A4 protein expression and mRNA levels were significantly reduced in HeLa and SiHa cells with arctiin treatment. Furthermore, silencing S100A4 by using small interfering RNA reduced cell migration, while overexpression of S100A4 mitigated the migration inhibition imposed by arctiin in cervical cancer cells. Western blotting revealed that arctiin significantly reduced phosphoinositide 3-kinase (PI3K) and phosphorylation of Akt in cervical cancer cells. Moreover, selective Akt induction by an Akt activator, SC-79, reverted cervical cancer cell migration and S100A4 protein expression, which were reduced in response to arctiin. Taken together, these results suggest that arctiin inhibits cervical cancer cell migration and invasion through suppression of S100A4 and the PI3K/Akt pathway.

The risk of distant metastases in patients with gynecologic cancers after surgery: a population-based study.

The aim of the study was to determine the risk of distant metastases in patients with gynecologic cancers after surgery, including cervical, uterine and ovarian cancers. This is a retrospective study evaluating gynecologic cancer from 2009 to 2014 using population-based administrative datasets from the Health and Welfare Data Science Center (HWDC) and from The National Health Informatics Project (NHIP). A total of 1,464 gynecologic cancer patients, including 321 cervical cancer patients, 724 uterine cancer patients and 419 ovarian cancer patients, were analyzed retrospectively from 2009 to 2014. Among the cervical cancer patients, 173 (53.89%) received surgery only and 148 (46.11%) received surgery with radiotherapy /chemotherapy. Among the uterus cancer patients, 425(58.70%) received surgery only and 299 (41.3%) received surgery with radiotherapy /chemotherapy. Among the ovarian cancer patients, 81 (19.33%) received surgery only and 338 (80.67%) received surgery with radiotherapy/chemotherapy. Among patients with brain, liver or lung metastasis, cervical cancer patients have more cumulative metastasis-free survival than those ovarian cancer (p=0.0041). In analyzing liver metastasis based on primary cancer sites, cervical cancer patients and uterine cancer cases have more cumulative metastasis- free survival than those ovarian cancer (p<0.0001). In conclusion, ovarian cancer patients have higher risk of liver metastasis than cervical or uterine cancer. There were significantly different of pathological stage for cumulative metastasis-free survival among gynecologic cancer patients with brain or liver or lung metastasis. Pathological T stage remains the main predictive for distant metastasis of gynecologic cancer.

Metformin Potentiates the Anticancer Effect of Everolimus on Cervical Cancer In Vitro and In Vivo.

Cervical cancer is globally the fourth most common cancer in women. Metformin is a widely used drug for the treatment of type II diabetes and has been shown to possess important anticancer properties in cervical cancer. Everolimus is an mTOR inhibitor and is widely used to treat NETs, RCC, TSC, and breast cancers. The present study investigated the anticancer effects of metformin and everolimus in cervical cancer, when used alone or in combination. CaSki and C33A human cervical cancer cells were treated with different concentrations of everolimus alone or in combination with metformin. Cell viability was assessed using a CCK-8 assay. Cell apoptosis, cell-cycle, and mtROS analyses were conducted using flow cytometry. Target protein levels were analyzed by Western blotting. Related mechanisms were confirmed using appropriate inhibitors (z-VAD-fmk and BIRB796). The in vitro results were further confirmed in a xenograft tumor study. Both metformin and everolimus, when used alone, were moderately effective in inhibiting cell proliferation and inducing cell apoptosis of CaSki and C33A cells. When used in combination, these two drugs synergistically inhibited the growth of human cervical cancer cells and xenografts in nude mice, promoted sub-G1- and G0/G1-phase cell-cycle arrest, and enhanced mtROS production. The protein expressions of PI3K (p110α) and p-AKT were significantly downregulated, while P27, P21, p-p38, p-ERK, and p-JNK were upregulated following combined treatment. These results revealed that metformin potentiates the anticancer effect of everolimus on cervical cancer, and combination treatment with metformin and everolimus provides a novel therapeutic strategy for patients with cervical cancer.

Molecular and Cellular Mechanisms of Metformin in Cervical Cancer.

Cervical cancer is one of the major gynecologic malignancies worldwide. Treatment options include chemotherapy, surgical resection, radiotherapy, or a combination of these treatments; however, relapse and recurrence may occur, and the outcome may not be favorable. Metformin is an established, safe, well-tolerated drug used in the treatment of type 2 diabetes; it can be safely combined with other antidiabetic agents. Diabetes, possibly associated with an increased site-specific cancer risk, may relate to the progression or initiation of specific types of cancer. The potential effects of metformin in terms of cancer prevention and therapy have been widely studied, and a number of studies have indicated its potential role in cancer treatment. The most frequently proposed mechanism underlying the diabetes-cancer association is insulin resistance, which leads to secondary hyperinsulinemia; furthermore, insulin may exert mitogenic effects through the insulin-like growth factor 1 (IGF-1) receptor, and hyperglycemia may worsen carcinogenesis through the induction of oxidative stress. Evidence has suggested clinical benefits of metformin in the treatment of gynecologic cancers. Combining current anticancer drugs with metformin may increase their efficacy and diminish adverse drug reactions. Accumulating evidence is indicating that metformin exerts anticancer effects alone or in combination with other agents in cervical cancer in vitro and in vivo. Metformin might thus serve as an adjunct therapeutic agent for cervical cancer. Here, we reviewed the potential anticancer effects of metformin against cervical cancer and discussed possible underlying mechanisms.

Impact of pentraxin 3 genetic variants on uterine cervical cancer clinicopathologic characteristics.

The aims of this study were to investigate the relationships among pentraxin 3 (PTX3) genetic variants and development and clinicopathological characteristics of uterine cervical cancer, and patient survival in Taiwanese women. The study enrolled 125 patients with invasive cancer and 98 patients with precancerous lesions of uterine cervix, and 325 control women. PTX3 genetic variants rs2120243, rs3816527, rs2305619 and rs1840680 were selected and their genotypic distributions were determined by real-time polymerase chain reaction. Our results indicated that patients with genotype CC in PTX3 rs2120243 and genotype GG in rs1840680 had more chance to have adenocarcinoma but not squamous cell carcinoma, as compared to those with CA/AA and those with GA/AA, respectively. No other clinicopatholgical characteristics were associated with PTX3 genetic variants. In addition, PTX3 genetic variants were not associated with 5 years survival of cervical cancer patients. In conclusions, PTX3 genetic variants are not associated with carcinogenesis and clinicopathological variables of uterine cervix and patient survival in Taiwanese women. The only independent predictor for the 5 years survival is pelvic lymph node metastasis.

Carbonic Anhydrase IX Promotes Human Cervical Cancer Cell Motility by Regulating PFKFB4 Expression.

Carbonic anhydrase IX (CAIX) is a hypoxia-induced protein that is highly expressed in numerous human cancers. However, the molecular mechanisms involved in CAIX and human cervical cancer metastasis remain poorly understood. In this study, CAIX overexpression in SiHa cells increased cell migration and epithelial-to-mesenchymal transition (EMT). Silencing CAIX in the Caski cell line decreased the motility of cells and EMT. Furthermore, the RNA-sequencing analysis identified a target gene, bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB4), which is influenced by CAIX overexpression and knockdown. A positive correlation was found between CAIX expression and PFKFB4 levels in the cervical cancer of the TCGA database. Mechanistically, CAIX overexpression activated the phosphorylation of extracellular signal-regulated kinases (ERKs) to induce EMT and promote cell migration. In clinical results, human cervical cancer patients with CAIXhigh/PFKFB4high expression in the late stage had higher rates of lymph node metastasis and the shortest survival time. Our study found that CAIX overexpression increases PFKFB4 expression and EMT, promoting cervical cancer cell migration. CAIX could contribute to cervical cancer cell metastasis and its inhibition could be a cervical cancer treatment strategy.

Pitavastatin and metformin synergistically activate apoptosis and autophagy in pancreatic cancer cells.

Pancreatic cancer is the seventh leading cause of cancer-related deaths globally. Metformin is the standard first-line of treatment for hyperglycemia in Type 2 diabetes, whereas pitavastatin is a cholesterol-lowering drug used to prevent cardiovascular diseases. Both these agents evidently exert anticancer effects on pancreatic cancer; however, it remains unclear whether cotreatment using them has additive or synergistic anticancer effects on pancreatic cancer. Thus, we herein used the ASPC-1 and PANC-1 cells and treated them with metformin and/or pitavastatin. We performed the cell viability assay, transwell migration assay, and cell cycle analysis using flow cytometry. Western blotting was used to determine protein levels. We found that cotreatment with metformin (30 mM) and pitavastatin (10 µM) significantly reduced cell viability; caused G0/G1 cell cycle arrest; upregulated the expression levels of Bax, PCNA, cleaved PARP-1, cleaved caspase-3, LC3 II, and p27 Kip1 /p21Cip1 ; and inhibited cell migration. The combination index value for cell viability indicated a synergistic interaction between metformin and pitavastatin. Moreover, cotreating the cells with metformin (30 mM) and pitavastatin (10 µM) could preserve mitochondrial function, activate AMPK, and inhibit PI3K/mTOR than treatment with metformin or pitavastatin alone. These findings clearly indicated that metformin plus pitavastatin had a synergistic anticancer effect on pancreatic cancer cells, potentially caused due to the activation of AMPK and inhibition of PI3K/mTOR signaling. Altogether, our results provide that use of metformin plus pitavastatin maybe serve as a chemotherapeutic agent for human pancreatic cancer in future.