CD24 gene inhibition and TIMP-4 gene upregulation by Imperata cylindrica's root extract prevents metastasis of CaSki cells via inhibiting PI3K/Akt/snail signaling pathway and blocking EMT.

ETHNOPHARMACOLOGICAL RELEVANCE: Imperata cylindrica (L.) Raeusch (Gramineae) is a medicinal spice traditionally used in the treatment of hypertension and cancer. AIM OF THE STUDY: To assess the anti-metastatic potential of the methanol extract of I. cylindrica roots and determined its mechanisms of action. MATERIAL AND METHODS: The growth inhibition activity of I. cylindrica root extract in vitro and in vivo in human cervical cancer. The scratch assay and Boyden Chamber assay were used to determine the anti-migrative and anti-invasion actions of the plant extract. The whole-genome gene expression profiling using RNA-Seq was performed to determine the differentially expressed genes in CaSki cells after exposure to I. cylindrica to identify its targeted genes related to metastasis. Using protein analysis (western blotting) and gene expression analysis (RTqPCR), the targeted pathways of the key genes that were initially identified with RNA-Seq, were evaluated. RESULTS: I. cylindrica extract showed dose-dependent cytotoxicity in vitro and in vivo in mice bearing tumors. Furthermore, I. cylindrica root extract significantly inhibited cell migration and cell invasion. After the genome-wide transcriptome analysis, we found that important genes involved in cancer progression and metastasis of cervical cancer, that is, CD24 and TIMP-4 were significantly downregulated and upregulated, respectively. Moreover, I. cylindrica root extract significantly inhibited the PI3/AKT/Snail signaling pathway and blocked the EMT of CaSki cells. CONCLUSION: These findings provide an anti-metastatic mechanism of action of I. cylindrica root extract toward the human cervical cancer suggesting that this plant maybe developed into selective chemotherapy.

Cytotoxic effect of aqueous ethanolic extract of Luffa cylindrica leaves on cancer stem cells CD44+/24- in breast cancer patients with various molecular sub-types using tissue samples in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Luffa cylindrica (L.) M.Roem is a climbing plant its parts have been used as traditional medicine for the treatment of different types of diseases including diarrhea, inflammation, cancer and viral infections. The parts used include fruit, seeds and leaves. AIM OF THE STUDY: Our study aims to investigate the effect of the aqueous-ethanol extract of Luffa cylindrica leaves on breast cancer stem cells CD44+/24- and other cell sub-populations using clinical samples with different molecular sub-types of breast cancer in vitro. MATERIALS AND METHODS: Breast tissues were obtained from patients undergoing surgery for the removal of breast tumors after complete clinical and pathological investigations. Tissue samples were processed to cell suspensions and treated with the extract in the tissue culture laboratory. Percentages of cell sub-populations within tumors and viability were measured by flowcytometry using clusters of differentiation as cell markers. RESULTS: Our results revealed that there were decreases in the total cell viability, CD44+/24- and total CD24+ cell sub-populations percentages after treatment with the extract, this may be an important indication of using Luffa leaves extract in the treatment of breast cancer or in combination with the traditional treatments. CONCLUSION: Luffa cylindrica has proven to have anticancer activity on three different subtypes of breast cancer including luminal A, luminal B and Her2/neu enriched more over it has cytotoxic effect on both bulk tumor cells as well as cancer stem cells sub population CD44+/24- which possess high tumorigenic potency, these results were confirmed by measuring their viable number after treatment and sphere formation assay results.

Molecular iodine impairs chemoresistance mechanisms, enhances doxorubicin retention and induces downregulation of the CD44+/CD24+ and E-cadherin+/vimentin+ subpopulations in MCF-7 cells resistant to low doses of doxorubicin.

One of the most dreaded clinical events for an oncology patient is resistance to treatment. Chemoresistance is a complex phenomenon based on alterations in apoptosis, the cell cycle and drug metabolism, and it correlates with the cancer stem cell phenotype and/or epithelial-mesenchymal transition. Molecular iodine (I2) exerts an antitumor effect on different types of iodine-capturing neoplasms by its oxidant/antioxidant properties and formation of iodolipids. In the present study, wild-type breast carcinoma cells (MCF-7/W) were treated chronically with 10 nM doxorubicin (DOX) to establish a low-dose DOX-resistant mammary cancer model (MCF-7/D). MCF-7/D cells were established after 30 days of treatment when the culture showed a proliferation rate similar to that of MCF-7/W. These DOX-resistant cells also showed increases in p21, Bcl-2 and MDR-1 expression. Supplementation with 200 µM I2 exerted similar effects in both cell lines: it decreased the proliferation rate by ~40%, and I2 co-administration with DOX significantly increased the inhibitory effect (to ~60%) and also increased apoptosis (BAX/Bcl-2 index), principally by inhibiting Bcl-2 expression. The inhibition by I2 + DOX was also accompanied by impaired MDR-1 induction as well as by a significant increase in PPARγ expression. All of these changes could be attributed to enhanced DOX retention and differential down-selection of CD44+/CD24+ and E-cadherin+/vimentin+ subpopulations. I2 + DOX-selected cells showed a weak induction of xenografts in Foxn1nu/nu mice, indicating that the iodine supplements reversed the tumorogenic capacity of the MCF-7/D cells. In conclusion, I2 is able to reduce the drug resistance and invasive capacity of mammary cancer cells exposed to DOX and represents an anti-chemoresistance agent with clinical potential.

[Xinfeng Capsule increases peripheral blood BTLA expression of CD19(+) and CD24(+) B cells and relieves oxidative stress damage to improve cardiac function of patients with rheumatoid arthritis].

OBJECTIVE: To observe the changes in cardiac function, peripheral blood B and T lymphocyte attenuator (BTLA) and oxidative stress related indicators in rheumatoid arthritis(RA) patients, thus to explore the mechanism underlying the improving effect of Xinfeng Capsule (XFC) on cardiac function. METHODS: The study enrolled 100 RA patients and divided them randomly into 2 groups, XFC treatment group and leflunomide (LEF) control group (n=50 per group). The treatment lasted 30 days for one course. Other 40 healthy people from Medical Examination Center were included as a normal control (NC) group. Flow cytometry was used to detect the expression and activation level of BTLA, Westergren method was used to determine erythrocyte sedimentation rate (ESR), and automatic biochemical analyzer to examine high sensitivity C-reactive protein (hs-CRP) and rheumatoid factor (RF). ELISA method was performed to observe related cytokines (IL-1ß, IL-17, IL-35, IFN-γ) and markers of oxidative stress such as total antioxidant capacity (TAOC), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione (GSH). Echocardiography was utilized to survey cardiac function parameters including heart ejection fraction (EF%), stroke volume (SV%), fractional shortening (FS%), E peak velocities (E) and A peak velocities (A) of mitral valve flow, and the ratio of filling fraction of E and A (E/A). RESULTS: Compared with the NC group, EF%, E peak velocity, E/A were significantly reduced while A peak velocity increased in RA patients, and FS% was not found obviously different. In addition, IL-1ß, IL-17 and inflammatory indexes like ESR, CRP increased while BTLA, IL-35, IFN-γ decreased significantly; Serum ROS, MDA rose and SOD, GSH dropped significantly in RA patients. Correlation analysis showed that the cardiac function parameters EF, FS were negatively correlated with CD24⁺ cells and CD19⁺CD24⁺ cells, that E/A was positively correlated with BTLA, that A peak velocity was positively related to CD19⁺ cells, that EF was positively related to ROS, that SV was positively related with MDA, SOD, that E peak velocity was negatively correlated with TAOC. After drug intervention, XFC treatment group got 86% total effective rate. XFC obviously improved cardiac function regarding EF%, FS%, E peak, E/A and other parameters, increased serum SOD, GSH capacity, eliminated ROS, MDA, increased BTLA, IL-35, IFN-γ, and reduced IL-1ß, IL-17. Compared with LEF group, XFC had a better improving effect on DAS28 and cardiac function parameters. CONCLUSION: XFC can increase BTLA expression of CD19⁺ and CD24⁺ B cells and reduce B cell-mediated abnormal humoral immunity and oxidative stress damage, thus improving cardiac function and quality of life.

The HMGB1/RAGE inflammatory pathway promotes pancreatic tumor growth by regulating mitochondrial bioenergetics.

Tumor cells require increased adenosine triphosphate (ATP) to support anabolism and proliferation. The precise mechanisms regulating this process in tumor cells are unknown. Here, we show that the receptor for advanced glycation endproducts (RAGE) and one of its primary ligands, high-mobility group box 1 (HMGB1), are required for optimal mitochondrial function within tumors. We found that RAGE is present in the mitochondria of cultured tumor cells as well as primary tumors. RAGE and HMGB1 coordinately enhanced tumor cell mitochondrial complex I activity, ATP production, tumor cell proliferation and migration. Lack of RAGE or inhibition of HMGB1 release diminished ATP production and slowed tumor growth in vitro and in vivo. These findings link, for the first time, the HMGB1-RAGE pathway with changes in bioenergetics. Moreover, our observations provide a novel mechanism within the tumor microenvironment by which necrosis and inflammation promote tumor progression.

One stop screening for multiple cancers: the experience of an integrated cancer prevention center.

BACKGROUND: Cancer is a leading cause of mortality worldwide. Screening is a key strategy for reducing cancer morbidity and mortality. METHODS: We aimed to describe the experience of an integrated cancer prevention center in screening an asymptomatic population for the presence of neoplasia. One-thousand consecutive asymptomatic, apparently healthy adults, aged 20-80 years, were screened for early detection of 11 common cancers that account for 70-80% of cancer mortality. RESULTS: Malignant and benign lesions were found in 2.4% and 7.1% of the screenees, respectively. The most common malignant lesions were in the gastrointestinal tract and breast followed by gynecological and skin. The compliance rate for the different screening procedures was considerably higher than the actual screening rate in the general Israeli population - 78% compared to 60% for mammography (p<0.001) and 39% compared to 16% for colonoscopy (p<0.001). Advanced age, family history of cancer and certain lifestyle parameters were associated with increased risk. Moreover, polymorphisms in the APC and CD24 genes indicated high cancer risk. When two of the polymorphisms existed in an individual, the risk for a neoplastic lesion was extremely high (OR 2.3 [95% CI 0.94-5.9]). CONCLUSIONS: One stop shop screening for 11 common cancers in the setting of a multidisciplinary outpatient clinic is feasible and can detect cancer at an early stage.