Targeted eradication of gastric cancer stem cells by CD44 targeting USP22 small interfering RNA-loaded nanoliposomes.

AIM: USP22, a member of ubiquitin-specific proteases (USPs), is a well-defined protein that promotes poor prognosis, invasion and metastasis, and also participates in the maintenance of cancer stem cells. USP22 siRNA-loaded nanoliposomes conjugated with CD44 antibodies (USP22-NLs-CD44) were constructed to enhance the therapeutic effect of USP22 siRNA against gastric cancer stem cells. MATERIALS & METHODS: The targeting and therapeutic efficacies of USP22-NLs-CD44 against gastric cancer stem cells were evaluated. RESULTS & CONCLUSION: USP22-NLs-CD44 was demonstrated to be able to effectively deliver USP22 siRNA to CD44+ gastric cancer stem cells, achieving superior therapeutic effects against CD44+ gastric cancer stem cells than nontargeted nanoliposomes. USP22-NLs-CD44 may provide a novel approach to eradicate gastric cancer stem cells in the near future.

Influence of total flavonoids derived from Choerospondias axillaris folium on aconitine-induced antiarrhythmic action and hemodynamics in Wistar rats.

It is well known that various traditional Chinese medicines produce antiarrhythmic actions. The aims of this study were to examine whether total flavones derived from Choerospondias axillaris folium (TFCF) also produced antiarrhythmic effects using a rat model of aconitine-induced arrhythmia and to compare these observations with the effects of total flavones of Choerospondias axillaris fructus (TFC). Wistar rats were orally administered TFC (0.2 g/kg) or TFCF (0.1, 0.2, or 0.4 g/kg) daily for 7 d. Subsequently, aconitine iv at 25 µg/kg was used to induce arrhythmia in these animals. Control (C) physiological saline and positive verapamil rats were also administered orally. The starting times of ventricular ectopic beats (VE), ventricular tachycardia (VT), ventricular fibrillation (VF), and heart arrest (HA) were recorded. In comparison to C, TFCF and TFC significantly prolonged the starting time of VE, VT, VF, and HA induced by aconitine. With respect to hemodynamics, TFC and high-dose TFCF were effective in reducing HR without associated changes in BP in all groups. TFC and TFCF decreased left ventricular systolic pressure (LVSP) and maximal velocity rate of ventricular pressure (+dp/dt max and -dp/dt min) with no marked effect on left ventricular end diastolic pressure (LVEDP) and -dp/dtmin. Data demonstrated that TFCF and TFC were equally effective in diminishing the aconitine-mediated arrhythmias. In addition, TFCF and TFC produced a similar reduction in HR with no accompanying change in BP. These findings indicate that the TFCF- and TFC-induced alterations may be attributed to inhibition of ventricular contraction without altering ventricular diastolic function.

[Research on Multi-Spectral Target Recognition System Based on the Magneto-Optical Modulation].

The technology of target recognition based on characteristic multi-spectrum has many advantages, such as strong detection capability and discriminating capability of target species. But there are some problems, it requires that you obtain the background spectrum as a priori knowledge, and it requires that the change of background spectrum is small with time. Thereby its application of real-time object recognition is limited in the new environment, or the complex environment. Based on magneto-optical modulation and characteristic multi-spectrum the method is designed, and the target is identified without prior access to the background spectrum. In order to achieve the function of the target information in the one acquisition time for tested, compared to conventional methods in terms of target detection, it's adaptability is better than before on the battlefield, and it is of more practical significance. Meanwhile, the magneto-optical modulator is used to suppress the interference of stray light background, thereby improving the probability of target recognition. Since the magneto-optical modulation provides incremental iterative target spectral information, therefore, even if the unknown background spectrum or background spectrum change is large, it can significantly improve the recognition accuracy of information through an iterative target spectrum. Different test targets back shimmering light intensity and background intensity values were analyzed during experiments, results showed that three targets for linearly polarized reflectance modulation is significantly stronger than the background. And it was of great influence to visible imaging target identification when measured target used camouflage color, but the system of polarization modulation type can still recognize target well. On this basis, the target range within 0.5 km x 2 km multi-wavelength characteristics of the target species were identified. When using three characteristic wavelengths, the probability of target identification significantly reduced at around 2km, when using four or five characteristic wavelength position, the probability of target identification reach up to more than 95.0%. Meanwhile, in order to reduce the calculation and improve the real-time detection capability of the system, finally, four characteristic wavelengths was selected. So the system has a certain application value.

[Imaging origins and characteristics analysis of acute and chronic aspiration pneumonia].

OBJECTIVE: To discuss about the pathologic and imaging origins and characteristics of CT scaning and X-ray radiography for acute and chronic aspiration pneumonia. METHODS: Imaging data from 30 patients with aspiration pneumonia were retrospectively analyzed, CT scaning was performed in 27 patients, which PMVR reconstruction was performed in 21 cases;3 exammed by X-ray with 2 used by esophagography. RESULTS: Opaque bodies were detected in trachea by CT scaning in 12 patients.7 patients in acute phase rapidly developed into acute respiratory distress syndrome(ARDS). CT signs of 30 patients with acute and chronic aspiration pneumonia included: centrilobular nodules were detected in 2 cases with acute phase, 4 cases with subacute phase and 4 cases with chronic phase; the imaging of ground glass opacity were detected in 9 cases with acute phase, 2 cases with subacute phase and 3 cases with chronic phase; the imaging of bronchiectasis was detected in 8 cases with chronic phase, which mucilage embolism was detected in 3 of 8 cases; the imaging of atelectasis was detected in 6 cases with chronic phase; the imaging of sheeted consolidation was detected in 5 cases with chronic phase, 8 case with acute phase; the imaging of interstitial fibrosis was detected in 3 cases with chronic phase. Lesions of inferior lobe of right lung were detected in 9 cases with chronic phase, 4 cases with subacute phase, 11 case with acute phase;lesions of inferior lobe of left lung were detected in 6 cases with chronic phase and 3 cases with subacute group, 11 case with acute phase. CONCLUSION: The imaging features of acute and chronic aspiration pneumonia overlap with GGO and centrilobular nodules in every group. While the imaging features of atelectasis, bronchiectasis or mucilage embolism are found in chronic phase. The chest CT scaning may accurately evaluate the dynamic change of aspiration pneumonia.

Pathologic features and clinical treatment of sarcomatoid intrahepatic cholangiocarcinoma.

The current study examined sarcomatoid intrahepatic cholangiocarcinoma (S-iCCA). S-iCCA was a more aggressive subtype of intrahepatic cholangiocarcinoma (iCCA). Early detection and complete resection of tumors are very important. Reported here is a case of S-iCCA, and the diagnosis and treatment of S-iCCA are discussed. The patient underwent a tumor resection and was treated with chemotherapy and molecularly targeted drugs after surgery. The clinical pathologic features and treatment of S-iCCA are discussed based on the literature. An immunohistochemical examination revealed positivity for cytokeratin 7 (CK7), CK-pan, vimentin, and CK19 and negativity for hepatocyte paraffin 1 (HepPar-1) in sarcomatoid cells. This case suggests that the particular molecular characteristics of sarcomatoid cells have great clinical diagnostic value, and comprehensive treatment of S-iCCA based on surgery is described.

Solid breast lesions: clinical experience with US-guided diffuse optical tomography combined with conventional US.

PURPOSE: To prospectively investigate the clinical efficacy of using diffuse optical tomography (DOT) and ultrasonographic (US) localization with conventional US to differentiate malignant solid breast lesions from those that are benign. MATERIALS AND METHODS: The study was approved by the institutional review board and all patients provided written informed consent. One hundred two consecutive women (mean age, 43 years; range, 18-86 years) who were referred for open biopsy with 136 breast lesions underwent conventional US and DOT with US localization. Total hemoglobin concentration and oxygen saturation were measured for each breast lesion. Sensitivity, specificity, overall accuracy, positive predictive value, and negative predictive value were determined with surgical pathologic examination results as the verification standard. RESULTS: Of the 136 biopsied lesions, 54 were carcinomas and 82 were benign. The average total hemoglobin concentration in the malignant group was 223.3 µmol/L±55.8 (standard deviation), and the average hemoglobin concentration in the benign group was 122.5 µmol/L±80.6 (P=.005). When the maximum hemoglobin concentration of 137.8 µmol/L was used as the threshold value, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of DOT with US localization were 96.3%, 65.9%, 65.0%, 96.4%, and 76.5%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of conventional US were 96.3%, 92.6%, 89.7%, 97.4%, and 93.4%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of conventional US combined with DOT were 100%, 93.9%, 91.5%, 100%, and 96.3%, respectively. CONCLUSION: US-guided DOT combined with conventional US improves accuracy compared with DOT alone.

[Based on Moire interference accurate detection of the laser center wavelength].

To improve spectrum resolution of the traditional Fourier interferometer with the same size lens, was proposed based on orthogonal wedge Fourier interferometer system. The interferometer system gets the optical path difference by the prism of two mutually perpendicular, which can make the laser Interferences on the CCD Array. The detector use the area array CCD linear array CCD, and the system collected the interference fringes on the two-dimensional plane. On the basis of the spectrum distribution of orthogonal inclination Moire interferometer by calculating optical path difference function, the system made the splicing of interference fringes on the area array CCD and Moire transform, finally got the spectrum resolution. The results from the MATLAB simulation software shows that the Maximum optical path difference of the orthogonal inclination Moire interferometer can be generated up to 234 microm, which is higher than the traditional Fourier interferometer about one order of magnitude, so the spectrum resolution also increased by nearly 10 times theoretical. Experimental calibration of the spectrometer with a selection of LAB SPAKR 750A-type spectrometer, measurements for the center wavelength of 635 nm semiconductor laser, the results show basically the same center wavelength position. However, the spectrum detected by orthogonal inclination Moire interferometer system near the center wavelength is better than the traditional interferometer system.

Effect of PI3K/AKT/mTOR Signaling Pathway on Regulating and Controlling the Anti-Invasion and Metastasis of Hepatoma Cells by Bufalin.

BACKGROUND: Autophagy plays a "double-edged sword" in the process of tumorigenesis, development and metastasis. OBJECTIVE: In this study, we explored the effect of PI3K/AKT/mTOR autophagy-related signaling pathway on regulating and controlling the invasion and metastasis of liver cancer cells by Bufalin. METHODS: The cell counting, migration, adhesion and invasion assay were used to evaluate the effect of Bufalin on cell proliferation, invasion and metastasis. The protein expression of PI3K/AKT/ mTOR signaling pathway were detected by the Western Blotting technique. RESULTS: After inhibiting autophagy of HCC-LM3 cells, the inhibitory effect of Bufalin on adhesion, migration and invasion of HCC-LM3 cells was significantly enhanced. Synergistic inhibition was strongest when different autophagy inhibitors were combined with 3MA and CQ. After inhibiting autophagy, Bufalin significantly inhibited the protein expression of P-AKT, Cyclin D1, MMP- 2, MMP-9 and VEGF in HCC-LM3 cells. The protein expression of PTEN and E-Cadherin in HCC-LM3 cells was significantly increased. CONCLUSION: The present study shows that the anti-tumor effect of Bufalin mainly inhibit proliferation, extracellular matrix degradation and angiogenesis of HCC by influencing autophagy. These findings confirm the capability of Bufalin in inhibiting metastasis of HCC and in parallel to current patents, could be applied as a novel therapeutic strategy in the prevention of metastasis of HCC.

Comparative proteomics analysis of human gastric cancer.

AIM: To isolate and identify differentially expressed proteins between cancer and normal tissues of gastric cancer by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). METHODS: Soluble fraction proteins of gastric cancer tissues and paired normal tissues were separated by 2-DE. The differentially expressed proteins were selected and identified by MALDI-TOF-MS and database search. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. Twenty-three protein spots were excised from sliver staining gel and digested in gel by trypsin, in which fifteen protein spots were identified successfully. Among the identified proteins, there were ten over-expressed and five under-expressed proteins in stomach cancer tissues compared with normal tissues. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human gastric cancer tissue and paired normal tissue were established and optimized and certain differentially-expressed proteins were identified. The combined use of 2-DE and MS provides an effective approach to screen for potential tumor markers.

Present status and recent advances in living donor liver transplantation for malignant hepatic tumors.

BACKGROUND: Living donor liver transplantation (LDLT) has been increasingly used to treat hepatic tumors worldwide in recent years, and is currently the most effective alternative to deceased donor liver transplantation to overcome the problem of organ shortage. LDLT has played an enormous role in treating early malignant hepatic tumors. But the indication of LDLT for malignant hepatic tumors is based on indefinite criteria. This review summarizes the recent studies in LDLT for treating malignant hepatic tumors. DATA SOURCES: A literature research of the PubMed database was conducted and research articles were reviewed. RESULTS: The current data on LDLT for malignant hepatic tumors, combined with our hospital experience, indicated that if a patient with hepatocellular carcinoma (HCC) who meets with the conventional Milan criteria cannot undergo tumor resection because of poorly preserved liver function, and a cadaveric graft is difficult to obtain within six months, LDLT may be selected. In a patient with recurrence of HCC after conventional therapies, feasibility, optimal timing, and efficacy of LDLT as a second-line treatment should be determined. CONCLUSIONS: Tumor recurrence is related to the biological behavior and staging of the tumor. New immunosuppressors which have anti-tumor effects and inhibit the immune system need to be developed. The indications of LDLT for hepatic malignant tumors should be selected meticulously.

The biological role of autophagy in regulating and controlling the proliferation of liver cancer cells induced by bufalin.

Autophagy is an important mechanism which regulates the processes of cell growth and death. The biological role of autophagy in regulating and controlling the proliferation of liver cancer cells by bufalin remains unknown. In the present study we investigated the effect of bufalin on autophagy of liver cancer cells. The growth inhibition and autophagy of liver cancer cells were detected using acridine orange fluorescence staining, flow cytometry and transmission electron microscopy. Combined with autophagy inhibitors (3­MA and CQ), CCK8 staining and western blot analysis were used to detect the effect of bufalin on the proliferation and autophagy­related protein expression in HCC­LM3 cells at the indicated time­points. Results indicated that combined with autophagy inhibitors 3­MA and CQ, the inhibitive effect of bufalin on the proliferation of HCC­LM3 cells was significantly enhanced. When combined with autophagy inhibitors 3­MA or CQ, bufalin significantly reduced the autophagosome and acidic vesicles in HCC­LM3 cells. When combined with autophagy inhibitors 3­MA and/or CQ for 12 h, bufalin significantly inhibited the expression of LC3­I and Beclin­1 in HCC­LM3 cells, and upregulated the expression of LC3­II and P62 in HCC­LM3 cells. When combined with autophagy inhibitors 3­MA and/or CQ for 24 h, bufalin significantly inhibited the LC3­II expression in HCC­LM3 cells, and upregulated the LC3­I, P62 and Beclin­1 expression in HCC­LM3 cells. When combined with autophagy inhibitors 3­MA and/or CQ for 48 h, bufalin significantly inhibited the expression of LC3­II and Beclin­1 in HCC­LM3 cells, and upregulated the expression of LC3­I and P62 in HCC­LM3 cells. These findings indicated that bufalin induced cell autophagy and inhibited proliferation of liver cancer cells by influencing the expression of autophagy related proteins including LC3­I, LC3­II, P62 and Beclin­1 in liver cancer cells. The autophagic state of liver cancer cells affected the inhibitory effect of bufalin on the proliferation of liver cancer cells.

Identification of the Differential Expression Profiles of Serum and Tissue Proteins During Rat Hepatocarcinogenesis.

The pathogenesis of hepatocellular carcinoma is complex and not fully known yet. This study aims to screen and identify the differentially expressed proteins in peripheral blood and liver tissue samples from rat hepatocellular carcinoma and to further clarify the pathogenesis and discover the specific tumor markers and molecular targets of hepatocellular carcinoma. The hepatocellular carcinoma model of Wistar rats were induced by chemical carcinogen. The serum and liver tissue samples were obtained after induction for 2, 4, 8, 14, 18, and 21 weeks. The results showed that the clusterin (IPI00198667), heat shock protein a8 (IPI00208205), and N-myc downstream-regulated gene-2 (IPI00382069) being closely related to hepatocarcinogenesis were eventually identified from the 30 different proteins. As the time progressed, the serum levels of clusterin and heat shock protein a8 increased gradually during induced liver cancer in rats. However, the serum N-myc downstream-regulated gene 2 level in induced liver cancer in rats underwent biphasic changes, and the serum N-myc downstream-regulated gene 2 level decreased at the 8th week, increased at the 14th week, and then decreased significantly. Statistical difference occurred in protein expression of clusterin and heat shock protein a8 in liver tissues at the different time points. In the liver tissues, the N-myc downstream-regulated gene 2 level decreased gradually at the 8th week, increased gradually at the 14th week, and then decreased significantly after 14 weeks. The study demonstrated that heat shock protein a8, clusterin, and N-myc downstream-regulated gene 2 participated in the process of abnormal cell division, proliferation, and carcinogenesis of liver cells during hepatocarcinogenesis.

SATB1 siRNA-encapsulated immunoliposomes conjugated with CD44 antibodies target and eliminate gastric cancer-initiating cells.

Purpose: Gastric cancer, the cancer initiated from the stomach, is ranked as the third most frequent reason of cancer death worldwide. Gastric cancer-initiating cells (CICs) are one of the crucial causes for the metastasis and recurrence of gastric cancer, and CD44 is considered to be one marker for gastric CICs. Special AT-rich sequence binding protein 1 (SATB1) is a protein that promotes cancer progression, metastasis, and invasion and also participates in the maintenance of CICs. In this study, we investigated the therapeutic effect of SATB1 siRNA against gastric CICs and we constructed SATB1 siRNA-encapsulated immunoliposomes conjugated with CD44 antibodies (CD44-SATB1-ILs) to enhance the therapeutic effect of SATB1 siRNA against gastric CICs. Methods: We investigated the therapeutic effect of the SATB1 suppression by SATB1 siRNA on CD44+ gastric CICs. CD44-SATB1-ILs were developed by the lyophilization/hydration approach. The targeting and cytotoxic effect of CD44-SATB1-ILs toward gastric CICs were evaluated in vitro. Results: In this study, for the first time, we confirmed that SATB1 suppression by SATB1 siRNA preferentially eliminated CD44+ gastric CICs. The results showed that CD44-SATB1-ILs could efficiently and specifically promote the SATB1 siRNA delivery to CD44+ gastric CICs, achieving superior therapeutic effects against CD44+ gastric CICs than non-targeted liposomes. Conclusion: As far as we know, our report is the first research that indicated the promotion of siRNA delivery via nanoparticles to gastric CICs and achievement of superior therapeutic effect against gastric CICs by utilization of CD44 antibody. Therefore, CD44-SATB1-ILs represent an up-and-coming approach for eliminating gastric CICs and also a promising treatment for therapy of gastric cancer.

Antitumor effects of brucine immuno-nanoparticles on hepatocellular carcinoma in vivo.

In vitro and in vivo studies have demonstrated that brucine is able to inhibit the proliferation of liver cancer cells and growth of animal tumors, and may be a promising anticancer drug. However, high toxicity, poor water solubility, short half-life, narrow therapeutic window, and similar therapeutic and toxic doses limit its clinical application in the treatment of malignant tumors. In our previous study, brucine immuno-nanoparticles were successfully prepared and added to the culture medium of liver cancer SMMC-7721 cells, and the results indicated that the brucine immuno-nanoparticles were able to target the cell membrane of liver cancer SMMC-7721 cells and significantly inhibit the proliferation, adhesion, invasion and metastasis of SMMC-7721 cells. The aim of the present study was to investigate the antitumor effect of brucine immuno-nanoparticles in vivo by establishing an in situ transplanted liver cancer in nude mice. The results indicated that in vivo application of the brucine immuno-nanoparticles resulted in temporary liver and kidney damage, and significantly reduced the α-fetoprotein (AFP) secretion of tumor cells (Bru-NP-MAb vs. the other groups; P<0.05). The brucine concentration of tumor tissues in the brucine immuno-nanoparticles group was significantly increased compared with that of the brucine nanoparticles group (Bru-NP-MAb vs. Bru-NP group or brucine group; P<0.05). The brucine immuno-nanoparticles were able to inhibit tumor growth and cluster of differentiation 34 expression and angiogenesis of tumor tissues, and induce the apoptosis of tumor cells (Bru-NP-MAb vs. Bru-NP group or brucine group; P<0.05). In conclusion, as a novel type of targeted drug, brucine nanoparticles combined with anti-AFP monoclonal antibodies was more effective compared with brucine nanoparticles or brucine alone in inhibiting tumor growth via the enhancement of apoptosis, and the suppression of proliferation and angiogenesis in vivo. Therefore, the brucine immuno-nanoparticle is a promising targeted drug for the treatment of hepatocellular carcinoma.

Effect of Sirpalpha1 on the expression of nuclear factor-kappa B in hepatocellular carcinoma.

BACKGROUND: Signal regulatory protein alpha1 (Sirpalpha1) is a member of Sirps families containing four immunoreceptor tyrosine-based inhibitory motifs (ITIMs) domains in the cytoplasm of and an activated substrate of receptor tyrosine kinase (RTK), that negatively regulates the RTK-dependent cell proliferating signal transduction pathway. Previously we found that Sirpalpha1 was closely associated with the occurrence and development of hepatocellular carcinoma (HCC) as well as liver regeneration. Since it is unclear about the regulatory mechanisms, we established the cell line transfected Sirpalpha1 gene and preliminarily clarified the mechanisms by which Sirpalpha1 negatively regulates the carcinogenesis and development of HCC. METHODS: Liver cancer Sk-Hep1 cell was respectively transfected with plasmids of pLXSN, pLXSN-Sirpalpha1 and pLXSN-Sirpalpha1delta4Y2, screened with the drug of G418 (1200 microg/ml), and various transfected Sk-Hep1 cell lines were obtained. The protein expressions of P65, P50, IkappaBalpha, cyclin D1 and Fas in various Sk-Hep1 cell lines were determined by Western blotting, and P65 and P50 were localized by the immunofluorescence technique. RESULTS: Sirpalpha1 could significantly upregulate the protein expression of IkappaBalpha (vs. other cell lines, P<0.05) in the Sk-Hep1 cell, and downregulate the protein expressions of P65, P50 and cyclin D1 (vs. other cell lines, P<0.05) in the Sk-Hep1 cell. P65 protein expression was mainly localized in the cytoplasm in the pLXSN Sk-Hep1 cell, and in the nucleus of the Sk-Hep1 cell with mutant Sirpalpha1delta4Y2, but in nucleus of the Sk-Hep1 cell with wild Sirpalpha1. P50 protein expression was localized in the cytoplasm and nucleus of the pLXSN Sk-Hep1 cell, but in the nucleus of the Sk-Hep1 cell with wild Sirpalpha1 and mutant Sirpalpha1delta4Y2 plasmid. CONCLUSIONS: Sirpalpha1 might negatively regulate and control the abnormal proliferation of liver cancer cells by influencing the protein content and localization of nuclear factor-kappa B, then influence the expression of cyclins such as cyclin D1 in the signal transduction pathway. It may be one of the important mechanisms by which Sirpalpha1 negatively regulates the carcinogenesis and development of HCC.

Preparation, pharmacokinetics, tissue distribution and antitumor effect of sorafenib-incorporating nanoparticles in vivo.

Sorafenib is a molecularly targeted drug used for treating hepatocellular carcinoma. However, sorafenib may affect the function of normal hepatocytes, and the clinical application of sorafenib is limited due to its adverse effects. The aim of the current study was to improve the effectiveness of sorafenib by preparing it as a nanoparticle formulation using nanoprecipitation technology. Sorafenib was combined with a polyethylene glycol monomethyl ether-racemic polylactic acid copolymer. The properties of the nanoparticles, including particle size, ξ potential and release efficiency, were measured. The pharmacokinetic profile, tissue distribution and tumor-inhibiting effects of the nanoparticles were determined in vitro and in vivo. Compared with sorafenib, the nanoparticle formulation exhibited a significant increase in in vivo retention time. The concentration of sorafenib in tumor tissues was significantly higher than that in normal tissues following treatment with sorafenib nanoparticles. Sorafenib nanoparticles were more efficacious in inhibiting tumor growth compared with sorafenib alone. The results, provided they can be extended to humans, suggest that sorafenib nanoparticles may specifically target hepatocellular carcinoma.

Negatively regulating mechanism of Sirpalpha1 in hepatocellular carcinoma: an experimental study.

BACKGROUND: Signal regulatory protein (Sirp) is a recently isolated, cloned and identified inhibitor receptor distributed in the membrane of hematopoietic and nonhematopoietic cells. Sirp alpha1 (Sirpalpha1) is a member of Sirp families. Sirpalpha1 can bind SHP-2 in the form of tyrosine phosphorylation by SH2 effect and negatively regulate growth factor, oncogene, or insulin-induced responses as its substrate. This study aimed to preliminarily clarify the negatively regulating proliferation mechanism of Sirpalpha1 in liver cancer. METHODS: pLXSN, Sirpalpha1 and Sirpalpha1P4Y2 plasmids were respectively transfected into Sk-Hep1 liver cancer cell line, and various stable Sk-Hep1 cell lines were obtained with screening agent of G418 (1200 microg/ml). The expressing levels of cyclin D1, CDK4, Fas, beta-catenin and gankyrin in various cell lines were determined with Western blotting. Cell cycles were determined at 0, 12 and 24 hours with flow cytometry after various synchronous cell lines were cultured without serum for 72. Cell apoptosis induced with agent of TNF-alpha (50 ng/ml) was determined with flow cytometry at 0, 0.5, 1, 3, 6 and 12 hours. RESULTS: Sirpalpha1 could significantly decrease the expression of cyclin D1, beta-catenin and gankyrin, but it couldn't affect the expression level of CDK4 and Fas. When synchronous cells were cultured for 12 hours, S phase Sk-Hep1 cell transfected with Sirpalpha1 plasmid was the lowest [(31.92+/-0.22)% vs. other cell lines, P<0.05], and the cell line was highly sensitive to TNF-alpha agent for 1 hour. (59.31+/-0.59)% of apoptotic cells occurred (vs. the other time points, P<0.05). CONCLUSIONS: Sirpalpha1 might block the cell cycle of liver cancer, inhibit cell proliferation, promote cell apoptosis by decreasing the expression of cyclin D1, beta-catenin and gankyrin. It is one of the important mechanisms inhibiting the occurrence and development of hepatocellular carcinoma.

Inhibitory effect of bufalin combined with Hedgehog signaling pathway inhibitors on proliferation and invasion and metastasis of liver cancer cells.

Hepatocellular carcinoma (HCC) is difficult to diagnose early, resulting in only 30% resection rate. HCC is a relatively chemo-resistant tumor, molecular targeted therapy can only benefit approximately 30% patients with liver cancer. Bufalin (Bu) is one of the topoisomerase II inhibitors, many studies have recently focused on the anticancer activities of bufalin. In the present study, we report that bufalin can inhibit the proliferation, invasion and metastasis of liver cancer cells via the Hh signaling pathway. The human high metastasis potential LM3 hepatoma cells (HCC-LM3) were cultured in vitro, bufalin and/or Hedgehog signaling pathway inhibitors (GANT61, cyclopamine) was added into cell culture fluid for 72 h to observe the antitumor effect of bufalin. The results showed that bufalin was able to inhibit epithelial mesenchymal transition (EMT), and extracellular matrix (ECM) degradation and angiogenesis of liver cancer cells by influencing the expression of Ptch1'Gli1'Gli3 proteins in Hh signaling pathway. Bufalin could downregulate the downstream target molecules of MMP-2, MMP-9, ß-catenin and VEGF in liver cancer cells by influencing the Gli1 and Gli3 expression of Hh signaling pathway, and upregulate the E-cadherin expression of liver cancer cells by influencing the Gli3 expression of Hh signaling pathway. Therefore, the present study shows that bufalin combined with Hedgehog signaling pathway inhibitors can significantly reduce the malignant biological behavior of the liver cancer cells.

Effects of signal regulatory proteinalpha1 on proliferation of hepatocellular carcinoma: a preliminary study.

BACKGROUND: Signal regulatory protein alpha1 (Sirpalpha1) is a negative regulatory factor, and inhibits receptor tyrosine kinase-dependent cell proliferating signal. This study was undertaken to observe the effect of signal regulatory proteinalpha1 (Sirpalpha1)on gankyrin,cyclin D1,CDK4 and Fas expression in Sk-hep1 mouse hepatoma carcinoma cell line. METHODS: BOSC 23 packed cells were respectively transfected by means of recombinated retrovirus including pLXSN, pLXSN- Sirpalpha1 and pLXSN- Sirpalpha1P4Y2 with lipofectin, and various plasmid virus media(viral titer 2.1X10(6) CFU/ml) were collected and infected respectively in 80% confluent Sk-hep1 cells. Transfected Sk-hep1 cells were selectively screened with G418 (1200 mug/ml), and Sk-hep1 cell lines transfected with various plasmids were obtained. The protein expressions of gankyrin, cyclin D1, CDK4 and Fas in various Sk-hep1 lines were determined by Western blotting. Various Sk-hep1 lines were recovered to culture with 10% fetal bovine serum at 12 hours and 24 hours after starving culture with free serum for 72 hours, and cells were collected to determine the percentage of S phase cells of proliferating cycle by flow cytometry. RESULTS: Sirpalpha1 transfection remarkably downregulated gankyrin and cyclin D1 expression. Sirpalpha1P4Y2 downregulation of gankyrin expression was greater than that of Sirpalpha1 (P<0.05), but no significant effect of Sirpalpha1 and Sirpalpha1P4Y2 on CDK4 and Fas protein expression was observed in transfected Sk-hep1 lines (P>0.05). The percentage of S phase cells significantly decreased in Sk-hep1 cells transfected with Sirpalpha1 and Sirpalpha1P4Y2 plasmids (vs pLXSN Sk-hep1, P<0.05). The percentage of S phase cells in various Sk-hep1 cells increased when recovering to culture with 10% fetal bovine serum at 12 hours, but the percentage of S phase cells in Sk-hep1 cells transfected with Sirpalpha1 was the lowest (vs pLXSN and Sirpalpha1P4Y2 Sk-hep1, P<0.05). The percentage of S phase cells in transfected pLSXN Sk-hep1 cells was the largest (vs Sirpalpha1 and Sirpalpha1P4Y2 Sk-hep1, P<0.05). There was no significant difference between the transfected Sirpalpha1 Sk-hep1 cells and Sirpalpha1P4Y2 Sk-hep1 cells (P>0.05). CONCLUSIONS: Sirpalpha1 decreases gankyrin and cyclin D1 expression, and inhibits proliferation of liver carcinoma cells. It may be one of the forms for an Sirpalpha1 negative regulation of carcinogenesis and development of hepatocellular carcinoma.

[Comparing the actions of the three flavone ingredients in choerospondias axillaris on arrhythmias induced by aconitine].

OBJECTIVE: To compare the actions of the three flavone ingredients in choerospondias axillaris on arrhythmias Induced by aconitine. METHOD: Langendorff perfuse was applied in the experiment, the antiarrhythmic action was to study by using aconitine on the the isolated heart; The antiarrhythmic action of the three flavone ingredients in choerospondias axillaris was to study by using i.v. aconitine in rat to induce arrhythmias. RESULT: Compared with the NS group, sample 1 and sample 2 both significantly prolonged the beginning time of VF of isolated heart and increased the dosage of aconitine, sample 3 reduced the beginning time of VF of isolated heart and decreased the dosage of aconitine, sample 1 and sample 2 both greatly prolonged the beginning time of VE, VT, VF, HA; sample 3 greatly reduced the beginning time of VT,VF. The actions of the three samples were in a concentration-dependent way. CONCLUSION: Sample 1 and sample 2 both resisted the occurrence of arrhythmias induced by aconitine, sample 3 markedly promoted the occurrence of arrhythmias induced by aconitine.