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Yan Gu

Publications and source records attributed to Yan Gu.

At least 37 records · Page 2Linked to original sources
PubMed2005

Recurrent acute pancreatitis and its relative factors.

AIM: To evaluate the causes and the relative factors of recurrent acute pancreatitis. METHODS: From 1997 to 2000, acute pancreatitis relapsed in 77 of 245 acute pancreatitis patients. By reviewing the clinical treatment results and the follow-up data, we analyzed the recurrent factors of acute pancreatitis using univariate analysis and multivariate analysis. RESULTS: Of the 245 acute pancreatitis patients, 77 were patients with recurrent acute pancreatitis. Of them, 56 patients relapsed two times, 19 relapsed three times, each patient relapsed three and four times. Forty-seven patients relapsed in hospital and the other 30 patients relapsed after discharge. Eighteen patients relapsed in 1 year, eight relapsed in 1-3 years, and four relapsed after 3 years. There were 48 cases of biliary pancreatitis, 3 of alcohol pancreatitis, 5 of hyperlipidemia pancreatitis, 21 of idiopathic pancreatitis. Univariate analysis showed that the patients with local complications of pancreas, obstructive jaundice and hepatic function injury were easy to recur during the treatment period of acute pancreatitis (P=0.022<0.05, P=0.012<0.05 and P=0.002<0.05, respectively). Multivariate analysis showed that there was no single factor related to recurrence. Of the 47 patients who had recurrence in hospital, 16 had recurrence in a fast period, 31 after refeeding. CONCLUSION: Acute pancreatitis is easy to recur even during treatment. The factors such as changes of pancreas structure and uncontrolled systemic inflammatory reaction are responsible for the recurrence of acute pancreatitis. Early refeeding increases the recurrence of acute pancreatitis. Defining the etiology is essential for reducing the recurrence of acute pancreatitis.

Acute Disease↗
PubMed2005

Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens.

Two serine proteinase genes were isolated from Culex pipiens pallens as significantly up-regulated genes in a deltamethrin-resistant strain through a combination of suppression substractive hybridization and gene expression profiling by macroarrays. These two genes were found to be expressed at least threefold higher in the resistant strain than in the susceptible one. By using rapid amplification of cDNA ends to screen the constructed cDNA library, we cloned these two sequences. There were 909 bp with an open reading frame of 786 bp in the sequence of trypsin cDNA (GenBank/NCBI AF468495), the deduced protein had 261 amino acids, which was most similar to the trypsin gene of Anopheles gambiae. There were 992 bp with an open reading frame of 816 bp in the chymotrypsin cDNA (GenBank/NCBI AY034060), and its deduced amino acid sequence had 271 amino acids, which was most similar to the chymotrypsin-like protein from Aedes aegypti. The two genes were stably expressed in mosquito C6/36 cells, and the expected 29 and 30 kDa bands were shown with Western blot, respectively. In these cells, after deltamethrin treatment, they had protective effects on the viability. The results indicate that trypsin and chymotrypsin were more highly expressed in the deltamethrin-resistant strain, and was related to insecticide resistance in mosquitoes, Cx. pipiens pallens.

Amino Acid Sequence↗
PubMed2005

Connexin interaction patterns in keratinocytes revealed morphologically and by FRET analysis.

Multiple connexins, the major proteins of gap junctions, have overlapping expression in the human epidermis and are postulated to have a key role in keratinocyte differentiation and homeostasis. The functional importance of connexins in the epidermis is emphasised by the association of mutations in four human connexins with various hyperproliferative skin disorders. As immunohistochemistry demonstrated overlapping expression of specific connexins in keratinocytes, we performed colocalisation analyses and applied a modified FRET methodology to assess possible heteromeric interactions between different combinations of four wild-type (wt) and mutant connexins. The data generated indicate that there is evidence for multiple connexin interactions at the plasma membrane between (wt)Cx26, (wt)Cx30 and (wt)Cx31 in keratinocytes and thus, the potential for the formation of a large number of different channel types each with different channel properties. In addition, we demonstrate that the inherent in vitro trafficking defect of the skin disease mutations (D50N)Cx26 and (G11R)Cx30 can be overcome partially by the coexpression of different wild-type connexins but this rescue does not result in large gap junction aggregates at the plasma membrane. These data indicate that skin disease associated Cx26 or Cx30 mutations are likely to disrupt a number of different channel types important in distinct aspects of keratinocyte biology.

Connexin 26↗
PubMed2005

Anatomic variations of the hook of hamate and the relationship to carpal tunnel syndrome.

PURPOSE: To determine the incidence of anatomic variations of the hook of hamate and to evaluate its association with the development of carpal tunnel syndrome (CTS). METHODS: Radiographs of 3,218 hands (2,070 patients) were evaluated. This group included 2,866 hands diagnosed with CTS and 352 hands without the diagnosis of CTS. Using the carpal tunnel view variations of the hook of hamate were identified. Variants were classified based on appearance and measurements. Patients having variant hooks of hamate who were diagnosed with CTS then were compared with patients with variant hooks of hamate who had no evidence of CTS. Endoscopic carpal tunnel release using the Chow technique was performed on all patients with CTS who had a known variation of the hook of hamate without difficulty or complication. RESULTS: Variations of the hook of hamate were found in 96 hands: 42, bipartite hook; 50, hypoplastic hook; and 4, aplastic hook. Ninety-three hands with variant hooks of hamate were diagnosed with CTS. The remaining 3 hands identified with variants had no evidence of CTS. After comparing the 2 groups, we found that the incidence of a variant hook of hamate in the group with CTS was significantly greater than the incidence of variations in the group without CTS. CONCLUSIONS: Three variations of the hook of hamate were identified with radiographic evaluation using the carpal tunnel view. Variations are more prevalent than once thought. There was a significant increase in the incidence of variation in the hook of hamate in the group with CTS compared with the group without CTS.

Adolescent↗
PubMed2005

Effects of extracellular delta-aminolaevulinic acid on sodium currents in acutely isolated rat hippocampal CA1 neurons.

The effects of delta-aminolaevulinic acid (ALA) on voltage-gated sodium channel (VGSC) currents (I(Na)) in acutely isolated hippocampal CA1 neurons from 10- to 12-day-old Wistar rats were examined by using the whole-cell patch-clamp technique under voltage-clamp conditions. ALA from 0.01 microm to 20 microm was applied to the recorded neurons. Low concentrations of ALA (0.01-1.0 microM) increased I(Na) amplitude, whereas high concentrations of ALA (5.0-20.0 microM) decreased it. The average I(Na) amplitude reached a maximum of 117.4 +/- 3.9% (n = 9, P < 0.05) with 0.1 microM ALA, and decreased to 78.1 +/- 3.8% (n = 13, P < 0.05) with 10 microm ALA. ALA shifted the steady-state activation and inactivation curves of I(Na) in the hyperpolarizing direction with different V0.5, suggesting that ALA could depress the opening threshold of the voltage-gated sodium channel (VGSC) and thus increase the excitability of neurons through facilitating the opening of VGSC. The time course of recovery from inactivation was significantly prolonged at both low and high concentrations of ALA, whereas either low or high concentrations of ALA had no significant effect on the attenuation of I(Na) during stimulation at 5 Hz, indicating that the effect of ALA on VGSC is state-independent. Furthermore, we found that application of ascorbic acid, which blocks pro-oxidative effects in neurons, could prevent the increase of I(Na) amplitude at low concentrations of ALA. Baclofen, an agonist of GABAb receptors, induced some similar effects to ALA on VGSC, whereas bicuculline, an antagonist of GABAa receptors, could not prevent ALA-induced effects on VGSC. These results suggested that ALA regulated VGSC mainly through its pro-oxidative effects and GABAb receptor-mediated effects.

Aminolevulinic Acid↗
PubMed2005

Cloning and overexpression of CYP6F1, a cytochrome P450 gene, from deltamethrin-resistant Culex pipiens pallens.

CYP6F1 (GenBank/EMBL accession No. AY662654), a novel gene with a complete encoding sequence in the cytochrome P450 family 6, was cloned and sequenced from deltamethrin-resistant 4th instar larvae of Culex pipiens pallens. The cDNA sequence of CYP6F1 has an open reading frame of 1527 bp, which encodes a putative protein of 508 amino acid residues. The deduced amino acid sequence of CYP6F1 indicated that the encoded P450 has conserved domains of a putative membrane-anchoring signal, putative reductase-binding sites, a typical heme-binding site, an ETLR motif and substrate recognition sites. Semi-quantitative RT-PCR analysis indicated that the CYP6F1 gene was expressed to a greater extent in the deltamethrin-resistant strain than in the susceptible strain of Cx. pipiens pallens. The expression levels of the CYP6F1 gene in the deltamethrin-resistant 1st, 2nd, 3rd, 4th instar larvae and adult female mosquitoes differed, with highest expression levels in the 4th instar larvae. In addition, the CYP6F1 gene was stably expressed in mosquito C6/36 cells, and the expected 61.2 kDa band was identified by Western blotting. The cells transfected with CYP6F1 had an increased resistance to deltamethrin as compared with control cells. These results indicate that CYP6F1 is expressed at higher levels in the deltamethrin-resistant strain, and may confer some insecticide resistance in Cx. pipiens pallens.

Amino Acid Sequence↗
PubMed2005

[Detection of IFN-gamma level in single CD8+ T cell].

AIM: To establish a new method capillary electrophoretic immunoassay laser-induced fluorescence (CEIA-LIF), to detect IFN-gamma level in single CD8(+) T cell. METHODS: CD8(+) T cells were isolated from peripheral blood of two patients with severe aplastic anemia(SAA) and one normal person by Ficoll-Hypaque gradient centrifugation and then were purified by immunomagnetic microbead separation. Then purified CD8(+) T cells were incubated with digitonin for 15 min followed by FITC-anti-IFN-gamma mAb for 20 min. The single cell was detected continuously by CEIA-LIF. The feasibility of the method was confirmed by inverted microscopy and laser scanning confocal fluorescence microscopy. RESULTS: The IFN-gamma content in purified CD8(+) T cells was detected under the condition of cell-membrane integrity. The IFN-gamma level in single CD8(+) T cell from 2 SAA patients was (151.53+/-28.92)zmol and (223.72+/-45.23)zmol, respectively, and much higher that from normal control (47.47+/-17.97)zmol ( P=0.001). CONCLUSION: It is feasible to quantitate IFN-gamma in single CD8(+) T cell by CEIA-LIF. CEIA-LIF might be useful in the clinical detection of intracellular cytokines.

Antigen-Antibody Complex↗
PubMed2005

Prediction of treatment outcome following correction of anterior crossbites in the mixed dentition: orthodontic versus orthopaedic methods.

AIM: To separate subjects with anterior crossbite and mild maxillary deficiency into two groups: those that can be treated successfully with a 2 x 4 appliance and those that should be treated with reverse headgear and maxillary expansion. METHODS: Thirty mixed dentition subjects each with an anterior crossbite, a concave profile, a deficient maxilla, and an ANB angle between -2 and +1 degrees were used. Fifteen subjects (8 boys, 7 girls; Mean age: 9.17 +/- 1.1 years) were successfully treated with a 2 x 4 appliance (orthodontic treatment group) and remainder (5 boys, 10 girls; Mean age: 8.6 +/- 1.5 years) were successfully treated with reverse headgear and maxillary expansion after initial treatment with a 2 x 4 appliance had failed (orthopaedic treatment group). Unstandardised discriminant function coefficients and a constant were calculated for selected pretreatment cephalometric variables. RESULTS: The NSAr angle was significantly smaller (p < 0.01) and both anterior and posterior cranial base lengths were significantly longer in the orthodontic treatment group (p < 0.001) compared with the orthopaedic group. The prediction index developed was: (PI) = -0.2 NSAr + 0.243 S-N + 0.187 S-Ar + 3.366. The critical score was 0. With positive scores a 2 x 4 appliance can be used and with negative scores reverse headgear and expansion are indicated. CONCLUSION: An equation able to predict treatment success with either a 2 x 4 appliance or reverse headgear has been developed.

Algorithms↗
PubMed2004

Identification of the region in Cdc42 that confers the binding specificity to activated Cdc42-associated kinase.

The Rho family small G-protein Cdc42 has been implicated in a diversity of biological functions. Multiple downstream effectors have been identified. How Cdc42 discriminates the interaction with its multiple downstream effectors is not known. Activated Cdc42-associated tyrosine kinase (ACK) is a very specific effector of Cdc42. To delineate the Cdc42 signaling pathway mediated by ACK, we set about to identify the specific ACK-binding region in Cdc42. We utilized TC10, another member of the Rho family of G-proteins that is 66.7% identical to Cdc42, to construct TC10/Cdc42 chimeras for screening the specific ACK-binding region in Cdc42. A region between switch I and switch II has been identified as the specific ACK-binding (AB) region. The replacement of the AB region with the corresponding region in TC10 resulted in the complete loss of ACK-binding ability but did not affect the binding to WASP, suggesting that the AB region confers the binding specificity to ACK. On the other hand, replacement of the corresponding region of TC10 with the AB region enabled TC10 to acquire ACK-binding ability. Eight residues are different between the AB region and the corresponding region of TC10. The mutational analysis indicated that all eight residues contribute to the binding to ACK2. The assays for the Cdc42-mediated activation of ACK2 indicated that the AB region is essential for Cdc42 to activate ACK2 in cells. Thus, our studies have defined a specific ACK-binding region in Cdc42 and have provided a molecular basis for generating ACK binding-defective mutants of Cdc42 to delineate ACK-mediated signaling pathway.

Amino Acid Sequence↗
PubMed2004

Effect of 4-aminopyridine on synaptic transmission in rat hippocampal slices.

Extracellular field excitatory postsynaptic potentials (fEPSPs) were recorded in area CA1 of rat hippocampal slices in vitro. The responses evoked by spontaneously released glutamate and GABA were recorded from area CA1 pyramidal neurons in rat hippocampal slices in whole-cell mode. The glutamate and GABA receptor-associated ligand-gated currents were obtained from dissociated single hippocampal pyramidal cells. The results showed that 4-aminopyridine (4-AP) had obvious effects on both presynaptic and postsynaptic events. Applications of 4-AP in micromolar concentration resulted in persistent enhancement of the initial slope of fEPSPs with the half-maximal enhancement concentration (EC(50)) of 46.7+/-2.68 microM. At the concentration of 200 microM, 4-AP increased the initial slopes of the total fEPSPs, NMDA- and AMPA-mediated fEPSPs components to 225.6+/-23.8%, 177.4+/-20.1% and 142.3+/-18.9%, respectively, but had no effect on the fiber volley. The half-maximal stimulus intensity to induce responses was reduced from 5.14+/-0.27 to 3.58+/-0.23 V. The frequencies of mEPSCs and mIPSCs were increased to 324.2+/-25.4% and 287.3+/-36.3% by 200 microM 4-AP. The amplitude histograms of mEPSCs and mIPSCs were fitted with Gaussian distributions. After 200 microM 4-AP application, the first and second peaks in Gaussian distributions of mEPSCs were shifted from 8.73+/-0.94 and 17.78+/-2.13pA to 10.48+/-0.82 and 21.14+/-2.45 pA, while those of mIPSCs were shifted from 13.65+/-0.96 and 25.51+/-2.95 pA to 11.21+/-1.04 and 23.08+/-2.37 pA. At 200 microM, 4-AP reduced paired-pulse facilitation and accelerated synaptic fatigue induced by stimulation at 10 Hz (for 1 s) and the ratio of fEPSPs(10)/fEPSPs(1) was decreased from 1.62+/-0.16 to 0.61+/-0.15. At 200 microM, 4-AP inhibited postsynaptic GABA currents induced by 5 microM GABA to 68.2+/-15.5%: by countering the effect of enhanced release of GABA from presynaptic terminals, this could depress the inhibitory pathway. Also at 200 microM, 4-AP increased NMDA currents to 155.3+/-17.8%, but had no significant effect on AMPA currents (94.2+/-15.6%). Our experimental results thus show that 4-AP-induced changes of synaptic transmission in area CA1 of rat hippocampus may be attributed to 4-AP's effects on both presynaptic terminals and postsynaptic receptors.

4-Aminopyridine↗
PubMed2004

Material reaction to suture anchor.

The authors present a case of material reaction to suture anchor, which is rarely reported. Leukocyte rate, leukocyte differential, multiple cultures, and gram stain test could not prove infection. A second surgery for exploration of the shoulder joint was performed to reconstruct the rotator cuff without using anchors, and the rotator cuff tear healed after the second surgery. During the second surgery, the bone surrounding the anchors was found to be eroded and substituted with necrotic tissue. The anchors protruded outside the bone. The pathological examination of the necrotic tissue showed multinucleated giant cells of foreign body type, some of which had engulfed minute splinters of particulate foreign material. Either metal- or suture-induced bioreaction is suspected in this case.

Aged↗
PubMed2004

[Preliminary study of non-surgical treatment of severe Class III malocclusion in 18 patients of 12-20 years old].

OBJECTIVE: To analyze effects of non-surgical treatment on subjects of 12-20 years old with severe skeletal Class III deformity and to directly evaluate dental and facial profile changes. METHODS: Eighteen patients with severe skeletal Class III malocclusion (male 5, female 13), diagnosed as indication for orthognathic surgery, were included in the study. The average age was 14.6 +/- 2.5 years old., with age range from 12 years old to 20 years old. 12 cases were treated with Tip-Edge straight-wire technique and 6 cases with Begg light wire technique. The average treatment time was 2.5 +/- 0.8 years. The selection criteria included: (1) mesial or superior mesial Class III molar relationship, with maxillary first molar occlude the buccal groove of the mandibular second molars (2) no mandibular shift (3) ANB < -1.5 degrees (4) high angle cases with average value of SN-MP 34.9 degrees +/- 5.9 degrees, 4 cases present with openbite (5) concave facial profile (6) originally classified as surgery cases. Lateral cephalometric films taken at the beginning and the end of the treatment were analyzed with traditional cephalometric analysis. The arithmetic mean (Mean) and standard deviation (SD) were calculated for each variable. Paired t-test was performed to evaluate the significant treatment change. RESULTS: Normal overjet and overbite were established with proclination of upper incisors and retroclination of lower incisors. Inclination of upper incisors was increased 5.9 degrees when measured with the angle of upper incisor to SN plane (P < 0.01). Inclination of lower incisors was decreased 6.6 degrees when measured with the angle of lower incisor to mandibular plane (P < 0.001). Negative value of the distance difference of upper lip and lower lip to Sn-Pg' at the beginning of treatment changed to positive value with significant difference (P < 0.001). CONCLUSION: Successful treatment effects could be obtained with non-surgical therapy in severe skeletal Class III malocclusion in permanent dentition. Remarkable soft-tissue change was noted after the treatment and concave facial profile changed to straight profile.

Adolescent↗
PubMed2003

Beta-adrenergic receptor blockade modulates Bcl-X(S) expression and reduces apoptosis in failing myocardium.

The mechanisms by which beta-adrenergic receptor (beta-AR) blockade modulates apoptosis in heart failure (HF) are unclear. We examined the impact of beta-AR blockade with metoprolol on myocardial remodeling, apoptosis, pro-apoptotic (Fas, Fas ligand, Bax, and Bcl-X(S)) and anti-apoptotic (Bcl-X(L)and Bcl-2) gene expression, and Bcl-X(L) and Bcl-X(S) protein in post-infarction HF in rats. In untreated rats, there was significant (P < 0.001) LV dilatation and systolic dysfunction compared to sham. Myocardial apoptosis was significantly increased (P < 0.005). Fas, Bax, and Bcl-2 mRNA expression was unchanged. However, Fas ligand mRNA and Bcl-X(S) mRNA and protein, all undetectable in sham, were markedly elevated (P < 0.001), whereas Bcl-X(L) mRNA and protein was unchanged. Immunohistochemistry confirmed increased Bcl-X(S) staining in failing myocardium, with unchanged Bcl-X(L). Metoprolol treatment resulted in: (1) improved LV remodeling (P < 0.025), (2) reduced myocardial apoptosis (P < 0.005), and (3) selective reduction in myocardial Bcl-X(S) expression (P < 0.001) without change in Fas, Fas ligand, Bax, Bcl-2, or Bcl-X(L). Studies in isolated rat myocytes revealed that prolonged isoproterenol (ISO) stimulation significantly increased Bcl-X(S) protein, reducing the Bcl-X(L)/X(S) ratio and myocyte survival (P < 0.005). ISO-induced Bcl-X(S) expression was significantly attenuated (P < 0.001) by both metoprolol and CGP20712A, a beta1-AR selective antagonist, but not by ICI118,551, a beta2-AR selective antagonist. We conclude that adrenergic activation, such as occurs in HF, increases pro-apoptotic Bcl-X(S) expression via the beta1-AR. beta-AR blockade in HF reduces myocardial apoptosis; attenuation of Bcl-X(S) expression may be one mechanism underlying this effect.

Adrenergic beta-Antagonists↗
PubMed2003

Crystal structure of an inactive Akt2 kinase domain.

Akt/PKB represents a subfamily of three isoforms from the AGC serine/threonine kinase family. Amplification of Akt activity has been implicated in diseases that involve inappropriate cell survival, including a number of human malignancies. The structure of an inactive and unliganded Akt2 kinase domain reveals several features that distinguish it from other kinases. Most of the alpha helix C is disordered. The activation loop in this structure adopts a conformation that appears to sterically hinder the binding of both ATP and peptide substrate. In addition, an intramolecular disulfide bond is observed between two cysteines in the activation loop. Residues within the linker region between the N- and C-terminal lobes also contribute to the inactive conformation by partially occupying the ATP binding site.

Adenosine Triphosphate↗
PubMed2003

Preliminary investigation of nonsurgical treatment of severe skeletal Class III malocclusion in the permanent dentition.

The purpose of the study was to analyze the effects of nonsurgical treatment on subjects with a severe skeletal Class III deformity and to directly evaluate dental and facial profile changes. Eighteen patients with severe skeletal Class III malocclusions (5 males, 13 females), diagnosed as indication for orthognathic surgery, were included in the study. The average age was 13.7 +/- 2.5 years. All the cases were treated with Tip-Edge straight-wire technique or Begg light wire technique. Lateral cephalometric films taken at the beginning and at the end of treatment were analyzed with the Pancherz analysis and traditional cephalometric analysis. The arithmetic mean and standard deviation were calculated for each variable and paired t-test was performed. A mean reduction of 6.5 mm in the overjet was noted (P < .001), with skeletal changes and dental changes contributing 20% and 80% to the overjet correction, respectively. The inclination of the upper incisors to the SN plane was increased 5.9 degrees (P < .01). The inclination of the lower incisors to the mandibular plane was decreased 6.6 degrees (P < .001). The difference between the distance of the upper lip and lower lip to Sn-Pg' at the beginning of treatment changed from a negative value to a positive value with a significant difference (P < .001). Successful treatment effects can be obtained with nonsurgical therapy in severe skeletal Class III malocclusions in the permanent dentition. A remarkable soft tissue change was noted after the treatment, and the concave facial profile changed to a straight profile (see two case reports).

Adolescent↗
PubMed2003

Cardiac toxic effects of trans-2-hexenal are mediated by induction of cardiomyocyte apoptotic pathways.

Aldehydes are ubiquitous pollutants with well-indicated but ill-defined cardiovascular toxicity. To investigate the direct toxic effects of environmental aldehyde exposure on the myocardium, 8-wk-old male ICR (Institute of Cancer Research) strain mice were gavage fed trans-2-hexenal (0.1, 1, 10, or 50 mg/kg/wk) or corn oil (vehicle) for 4 wk, during which cardiac function, myocardial morphology, cardiomyocyte apoptosis, and the cytochrome cmediated caspase activation apoptotic pathway were determined. Quantification by enzyme-linked immunosorbent assay (ELISA) revealed that aldehyde- protein adducts increase in mouse hearts following hexenal treatment, whereas echocardiographic analysis displayed a significant impairment of basal left-ventricular contractile function. Both histological analysis and TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) staining indicated condensed nuclei and a significant increase in cardiomyocyte apoptosis in these mice, but immunohistochemistry-based confocal microscope revealed no marked myofibril disarray. Release of cytochrome c from mitochondria into the cytosol, concomitant with activation of caspase-3 and -9, was also found in hexenal-treated groups. In addition, isolated cardiac mitochondria formed hexenal-protein adducts when treated with hexenal, providing indirect evidence that the cardiac mitochondrion is one of primary subcellular targets of aldehyde toxins. These findings suggest that trans-2-hexenal exposure results in direct cardiac toxicity through, at least in part, induction of mitochondrial cytochrome c release-mediated apoptosis in cardiomyocytes, indicating that the cardiac mitochondrion is one of principal subcellular targets of aldehyde toxins.

Actinin↗
PubMed2003

[Inhibitory effects of antisense focal adhesion kinase oligodeoxynucleotides on the invasion of Bel 7402 hepatocellular carcinoma cells].

OBJECTIVE: To study the inhibitory effects of antisense focal adhesion kinase (FAK) oligodeoxynucleotides (ODN) on the invasion of Bel 7402 cells, and investigate the mechanisms. METHODS: LipofecTAMINE-mediated antisense FAK ODN was transfected into Bel 7402 cells. Cell number and viability were evaluated every 24 hours by trypan blue dye exclusion. Cell attachment assay was carried out at intended time points in a microculture well pre-coated with fibronectin (FN). The invasive activity of tumor cells was assayed in a transwell cell culture chamber. Cell cycle and cell apoptosis analysis were performed with flow cytometry (FCM). RESULTS: The expression of p125FAK in the group treated with antisense FAK ODN (6.49%+/-0.10%) significantly decreased, compared with those in the group treated with sense FAK ODN (14.33%+/-1.88%) and control group (16.68%+/-1.62%), F=7.66, P<0.01. Antisense FAK ODN significantly inhibited the growth of Bel 7402 cells by 30%-60%, the attachment by 25%-55%, and the invasion, 15%-25%. The decreased expression of FAK in Bel 7402 cells caused a G2/M cell cycle arrest, and the cells at S phase decreased significantly. The occurrence of apoptosis detected by FCM increased significantly in the group treated with antisense FAK ODN. CONCLUSIONS: Inhibition of FAK expression significantly decreases the attachment between ECM and Bel 7402 cells, and the ability of Bel 7402 cells to invade the reconstituted basement membrane. In addition, FAK suppression significantly inhibits the proliferation of Bel 7402 cells in vitro, and increases their apoptosis.

Apoptosis↗