Search

The post-retrieval extinction paradigm, rooted in reconsolidation theory, holds promise for enhancing extinction learning and addressing anxiety and trauma-related disorders. This study investigates the impact of two reminder types, mild US-reminder (US-R) and CS-reminder (CS-R), along with a no-reminder extinction, on fear recovery prevention in a categorical fear conditioning paradigm. Scalp EEG recordings during reminder and extinction processes were conducted in a three-day design. Results show that the US-R group exhibits a distinctive extinction learning pattern, characterized by a slowed-down yet successful process and pronounced theta-alpha desynchronization (source-located in the prefrontal cortex) during CS processing, followed by enhanced synchronization (source-located in the anterior cingulate) after shock cancellation in extinction trials. These neural dynamics correlate with the subtle advantage of US-R in the Day 3 recovery test, presenting faster spontaneous recovery fading and generally lower fear reinstatement responses. Conversely, the CS reminder elicits CS-specific effects in later episodic tests. The unique neural features of the US-R group suggest a larger prediction error and subsequent effortful conflict learning processes, warranting further exploration., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Great progress has been made in identifying positive regulators that activate adipocyte thermogenesis, but negative regulatory signaling of thermogenesis remains poorly understood. Here, we found that cardiotrophin-like cytokine factor 1 (CLCF1) signaling led to loss of brown fat identity, which impaired thermogenic capacity. CLCF1 levels decreased during thermogenic stimulation but were considerably increased in obesity. Adipocyte-specific CLCF1 transgenic (CLCF1-ATG) mice showed impaired energy expenditure and severe cold intolerance. Elevated CLCF1 triggered whitening of brown adipose tissue by suppressing mitochondrial biogenesis. Mechanistically, CLCF1 bound and activated ciliary neurotrophic factor receptor (CNTFR) and augmented signal transducer and activator of transcription 3 (STAT3) signaling. STAT3 transcriptionally inhibited both peroxisome proliferator-activated receptor-γ coactivator (PGC) 1α and 1β, which thereafter restrained mitochondrial biogenesis in adipocytes. Inhibition of CNTFR or STAT3 could diminish the inhibitory effects of CLCF1 on mitochondrial biogenesis and thermogenesis. As a result, CLCF1-TG mice were predisposed to develop metabolic dysfunction even without external metabolic stress. Our findings revealed a brake signal on nonshivering thermogenesis and suggested that targeting this pathway could be used to restore brown fat activity and systemic metabolic homeostasis in obesity.

The ability of emotion regulation under stress is of crucial importance to psychosocial health. Yet, the dynamic function of stress hormones for the cognitive control of emotions over time via non-genomic and genomic cortisol effects remains to be elucidated. In this randomized, double-blind, placebo-controlled neuroimaging experiment, 105 participants (54 men, 51 women) received 20 mg hydrocortisone (cortisol) or a placebo either 30min (rapid, non-genomic cortisol effects) or 90min (slow, genomic cortisol effects) prior to a cognitive reappraisal task including different regulatory goals (i.e., downregulate vs. upregulate negative emotions). On the behavioral level, cortisol rapidly reduced and slowly enhanced emotional responsivity to negative pictures. However, only slow cortisol effects improved downregulation of negative emotions. On the neural level, cortisol rapidly enhanced, but slowly reduced amygdala and dorsolateral prefrontal activation as well as functional connectivity between both structures in the down- minus upregulate contrast. This interaction speaks for an effortful but ineffective regulation of negative emotions during rapid cortisol effects and improved emotion regulation capacities during slow cortisol effects. Taken together, these results indicate a functional shift of cortisol effects on emotion regulation processes over time which may foster successful adaptation to and recovery from stressful life events., Competing Interests: None., (© 2023 The Authors.)

While interpretation-bias modification (IBM) is an effective intervention for treating anxiety, it is not broadly used in clinical or daily practice. To this end, this study developed and tested a smartphone-based IBM application. We adopted the ambiguous situation paradigm as an intervention task in conjunction with robust training materials that broadly covered situations encountered in daily life. We recruited participants with high-trait anxiety and divided them into three groups: (1) positive training; (2) 50% positive-50% negative training; and (3) no-training control. The first two groups completed 28 days of smartphone-based training (IBM in positive cases), and all groups completed six rounds of assessments. The smartphone-based IBM training changed positive and negative endorsements and more specific measures of interpretation bias, thus reducing anxiety. The results also showed that changes in the number of negative interpretations played a mediating role in anxiety reduction. It is notable that the attrition rate was extremely low across the experiment. Our follow-up showed that positive gains persisted throughout the intervening period. Smartphone-based IBM can help individuals with anxiety shift negative biases, broaden their thoughts, enhance their information processing, and effectively target the clinical features of anxiety.

The activation of brown adipose tissues and beige adipose tissues can utilize more substrates, including glucose and fatty acids, regulate the energy balance of the whole body and improve metabolic diseases such as obesity and type Ⅱ diabetes. Elucidating the regulatory mechanisms underlying the thermogenic adipose program may provide excellent targets for therapeutics against metabolic diseases. The current studies have indicated that epigenetic modifications are vital for regulating differentiation and thermogenesis of adipose tissues. In this review, we summarize the recent progress of epigenetic modifications in adipose tissue development and thermogenesis from the aspects of DNA methylation, histone modification, chromatin remodeling, and non-coding RNAs in order to provide new ideas for further studying the activation of adipose tissues.

Uncertainty of threat has been linked to anxiety, but little is known about how neurophysiological responses change as uncertain threats approach and whether trait anxiety modulates these changes. The current study was designed to explore aspects of the dynamics of defensive motivation (startle reflex elicited by acoustic startle probes), attention (probe N100 component of event-related potentials elicited by acoustic startle probes), and autonomic nervous system activity (heart rate) when the approaching threat was certain or uncertain in a variant of the threat probability task. Behavioral results showed that high-trait anxious individuals reported higher levels of anxiety than low-trait anxious individuals only under the uncertain threat condition. Electromyographic data showed that high-trait anxious individuals tended to produce a more pronounced startle reflex, especially when the uncertain threat was proximal. This pattern was not observed in low-trait anxious individuals. By examining early attention engagement through probe N100, we observed a similar pattern in relation to defensive motivation. Moreover, under the uncertain threat condition, high-trait anxious individuals yielded a greater deceleration of heart rate than low-trait anxious individuals. These results indicate a distinct modulation effect of trait anxiety in the dynamics of defensive motivation, attention, and the autonomic nervous system during the anticipation of uncertain threats., (© 2022 Society for Psychophysiological Research.)

Brown and beige adipocytes dissipate energy in a nonshivering thermogenesis manner, exerting beneficial effects on metabolic homeostasis. CHCHD10 is a nuclear-encoded mitochondrial protein involved in cristae organization; however, its role in thermogenic adipocytes remains unknown. We identify CHCHD10 as a novel regulator for adipocyte thermogenesis. CHCHD10 is dramatically upregulated during thermogenic adipocyte activation by PPARγ-PGC1α and positively correlated with UCP1 expression in adipose tissues from humans and mice. We generated adipocyte-specific Chchd10 knockout mice (Chchd10-AKO) and found that depleting CHCHD10 leads to impaired UCP1-dependent thermogenesis and energy expenditure in the fasting state, with no effect in the fed state. Lipolysis in adipocytes is disrupted by CHCHD10 deficiency, while augmented lipolysis through ATGL overexpression recovers adipocyte thermogenesis in Chchd10-AKO mice. Consistently, overexpression of Chchd10 activates thermogenic adipocytes. Mechanistically, CHCHD10 deficiency results in the disorganization of mitochondrial cristae, leading to impairment of oxidative phosphorylation complex assembly in mitochondria, which in turn inhibits ATP generation. Decreased ATP results in downregulation of lipolysis by reducing nascent protein synthesis of ATGL, thereby suppressing adipocyte thermogenesis. As a result, Chchd10-AKO mice are prone to develop high-fat diet-induced metabolic disorders. Together, our findings reveal an essential role of CHCHD10 in regulating lipolysis and the thermogenic program in adipocytes., (© 2022 by the American Diabetes Association.)

Background: The effect of working memory training (WM-T) has been found to transfer to emotional wellbeing, despite some debate on whether an affective component in training is necessary to achieve specific emotion-related benefits. These novel cognitive trainings have not yet been tested in highly anxious individuals, who have deficits in implicit and explicit emotional regulation and should be the potential beneficiaries of these trainings., Methods: We designed two types of mobile phone-based training applications: (1) WMT and (2) an emotional working memory training (EWM-T) that comprised negative face distraction. Ninety-eight participants (33, WM-T; 35, EWM-T; 30, Control group) with high trait anxiety completed the 21-day intervention or placebo program and conducted pre- and post-test procedures, including questionnaires, emotional regulation and emotional Stroop tasks alongside electroencephalogram recording. Late positive potential (LPP) in emotion regulation task and P3 in the emotional Stroop task were adopted as neutral indicators for the explicit and implicit affective regulation/control processing., Results: Those who had received training (WM-T and EWM-T) showed enhanced explicit regulation (indexed by reduced LPP during reappraisal) compared with the control. Besides, individuals in EWM-T showed reduced behavioral attention bias and a decline of P3 in response to negative faces in an emotional Stroop task. The altered neural indicators were correlated with corresponding behavior indexes that contributed to the anxiety alleviation., Conclusions: The general WM-T was effective in enhancing explicit emotional regulation, while training with emotional add-in further improved implicit emotional control. (E)WM-T shows potential as a beneficial intervention for the anxiety population.

Individuals have a tendency to show enhanced vigilance to groups of which they themselves are not a member. Stress can up-regulate hypervigilance towards threatening stimuli and was shown to promote the reinstatement of out-group related biases in a previous study conducted in women only. The current study examines how exposure to acute stress affects the retrieval of out-group related extinction biases in male participants. Results showed that men exerted a specific out-group related bias at the beginning of extinction training indexed by higher skin conductance responses (SCRs) towards out-group faces, while stress led to a return of this extinguished out-group bias. Specifically, the stress group showed higher SCRs towards out-group faces during retrieval compared to the control group and the bias index was negatively related to post-stress cardiovascular recovery. These results indicate the important interaction between stress and intergroup bias in fear conditioning, along with a potential modulation of sex., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

When memories are reactivated, they enter a period of instability in which they can be affected by a variety of follow-up manipulations. The existence of this type of memory reconsolidation offers the potential for clinical interventions of maladaptive memory. However, such potential cannot be fully exploited until the internal mechanisms of memory changes via reconsolidation are better understood. In the current study, we used a three-day AB-AC paradigm that included self-referential simulation processing and employed electroencephalogram (EEG) techniques to explore how post-retrieval updates of episodic memory come about. Behaviorally, we found that reactivation alongside interference learning (ReI-L, AB-AC, n = 52) can produce much more false memories compared to no reactivation new learning (New-L, AB-DC, n = 31) and reactivation repetitive learning (Rep-L, AB-AB. n = 30). More importantly, ERP results revealed that trials from ReI-L in which memory distortions subsequently occurred showed an observable (compared to the new-learning without memory reactivation) but attenuated (compared to trials associating with later intact memory) amplitude of frontal N400, indicating a moderate level of early conflict reactivation is necessary to trigger crucial memory instability. In addition, to promote optimal distortion of the original memory, a sufficient later constructional processing is also required, reflecting in these intrusive/later false trials showed a larger amplitude of late posterior negativity (LPN). A linear classifier employing neuro features of FN400 and LPN during the reconsolidation phase could predict the original memory retention with 72% accuracy. The present findings indicate that nuance in post-retrieval interference, moderate conflict with protracted construction can lead to optimal alterations of episodic memories., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background and Aims: NAFLD, characterized by aberrant triglyceride accumulation in liver, affects the metabolic remodeling of hepatic and nonhepatic tissues by secreting altered hepatokines. Small ubiquitin-related modifier (SUMO)-specific protease 2 (SENP2) is responsible for de-SUMOylation of target protein, with broad effects on cell growth, signal transduction, and developmental processes. However, the role of SENP2 in hepatic metabolism remains unclear., Approach and Results: We found that SENP2 was the most dramatically increased SENP in the fatty liver and that its level was modulated by fed/fasted conditions. To define the role of hepatic SENP2 in metabolic regulation, we generated liver-specific SENP2 knockout (Senp2-LKO) mice. Senp2-LKO mice exhibited resistance to high-fat diet-induced hepatic steatosis and obesity. RNA-sequencing analysis showed that Senp2 deficiency up-regulated genes involved in fatty acid oxidation and down-regulated genes in lipogenesis in the liver. Additionally, ablation of hepatic SENP2 activated thermogenesis of adipose tissues. Improved energy homeostasis of both the liver and adipose tissues by SENP2 disruption prompted us to detect the hepatokines, with FGF21 identified as a key factor markedly elevated in Senp2-LKO mice that maintained metabolic homeostasis. Loss of FGF21 obviously reversed the positive effects of SENP2 deficiency on metabolism. Mechanistically, by screening transcriptional factors of FGF21, peroxisome proliferator-activated receptor alpha (PPARα) was defined as the mediator for SENP2 and FGF21. SENP2 interacted with PPARα and deSUMOylated it, thereby promoting ubiquitylation and subsequent degradation of PPARα, which in turn inhibited FGF21 expression and fatty acid oxidation. Consistently, SENP2 overexpression in liver facilitated development of metabolic disorders., Conclusions: Our finding demonstrated a key role of hepatic SENP2 in governing metabolic balance by regulating liver-adipose tissue crosstalk, linking the SUMOylation process to metabolic regulation., (© 2021 by the American Association for the Study of Liver Diseases.)

According to the theory of reconsolidation, the contents of an original memory can be updated after reactivation with subsequent new learnings. However, there seems to be a lack of an appropriate behavioral paradigm to study the reconsolidation of explicit self-related memory, which is of great significance to further explore its cognitive neural mechanism in the future. In two separate experiments, we adapted a trial-by-trial interfering paradigm with a self-episodic simulation process and investigated (1) whether it is possible to reconsolidate negative memories under the new behavioral paradigm and (2) how the emotional valence of post-retrieval interference material affects the reconsolidation of negative memories. The results showed that the negative memories under trial-by-trial self-simulation can be degraded and updated via post-retrieval interference processes. Individuals whose original memories were reactivated by initial background cues and who were then presented with new interference situations were less able to recall original scenes and showed more memory intrusions on these scenes than those who had experienced new learning without reactivation or only reactivation without interference. Furthermore, the extent and manner of memory change/updating were greatly influenced by the characteristics of interference information. For memories with negative valences, new learning materials with the same valence produced superior interference effects in the form of lower correct recalls and more integrated false; whereas the neutral interference materials can cause more memory intrusion. Post-retrieval memory distortions of negative self-memory may underlie different functional mechanisms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Pan and Li.)

Although impaired ability to regulate emotion is commonly reported in schizophrenic patients, the exact pattern of regulation of negative emotions in high-risk individuals remains unclear. In the current study, 26 high-schizotypy individuals paired with 26 controls completed an emotion regulation questionnaire (ERQ) and a laboratory emotion regulation task with electroencephalogram (EEG) recording. Two emotion regulation strategies, namely, reappraisal and expression suppression, were concurrently examined. The late positive potential (LPP) and frontal alpha asymmetry (FAA) were selected as two independent neural indicators of the emotion regulation effect. In the ERQ questionnaire, individuals in the high schizotypy group reported higher habitual use of suppression than the controls. During the emotion regulation task, the high  schizotypy group showed no early LPP reduction in reappraisal compared with the control group and exhibited a general negative FAA pattern (left-biased alpha). In conclusion, we found that individuals with high schizotypy exhibited maladaptive regulation of negative emotions, manifested in hindered reappraisal and biased suppression; this may exacerbate the negative affect of such emotions and further serve as a risk factor for psychosis conversion. Early interventions targeting the regulation of negative emotions may be beneficial for individuals with high schizotypal traits.

Schizotypy is associated with poor emotion regulation that is thought to contribute to the development of psychotic symptoms and to indicate a predisposition to schizophrenia. Having focused primarily on the relationship between schizotypy and explicit emotion regulation, existing studies have, until now, neglected to acknowledge the potentially important role of implicit emotion regulation. Our aim in the current study was to investigate implicit emotion regulation deficits in schizotypy. To this end, we used a newly developed Priming-Identification (PI) ERP paradigm, consisting of a priming phase and an emotion identification phase, to test 30 individuals with schizotypy and 30 healthy controls while also acquiring EEG data. During the priming phase, we aimed to manipulate emotion regulation goals (i.e., to bring about an intended emotional state) by presenting a category of words related to emotion regulation alongside a category of control words. Associated brain responses occurring during the subsequent stage were indexed according to three ERP components: N170, early posterior negativity (EPN) and late positive potential (LPP). Results showed that, in the control group, priming words associated with emotion regulation led to enhancements in the early N170 amplitude and the middle EPN during expression identification. The same pattern was not observed in the schizotypy group. In summary, our results suggest the presence of deficits in the early and middle stages of the implicit emotion regulation process among individuals with high schizotypal traits.

Rumination is an important etiological factor of anxiety pathology, with its mechanism related to the deficit of working memory. The current study examined whether working memory training (WM-T) and emotional working memory training (EWM-T) could reduce rumination in anxious individuals. The participants with high trait anxiety underwent 21 days of mobile applications-based WM-T (n = 34), EWM-T (n = 36) or placebo control (n = 36), with questionnaires, cognitive tasks, and resting electroencephalogram (EEG) as the pre-post-test indicators. The results revealed that two training groups obtained comparable operation span increases (WM-T: d = 0.53; EWM-T: d = 0.65), updating improvement (WM-T: d = 0.43; EWM-T: d = 0.60) and shifting improvement (WM-T: d = 0.49; EWM-T: d = 0.72). Furthermore, compared to the control group, the EWM-T showed significant self-reported rumination reduction (d = 0.69), increased inhibition ability (d = 0.72), as well as modification of resting EEG microstate C parameters (Duration C: d = 0.42, Coverage C: d = 0.39), which were closely related to rumination level (r ~ 0.4). The WM-T group also showed the potential to reduced self-reported rumination (d = 0.45), but with the absence of the observable inhibition improvement and resting EEG changes. The correlation analysis suggested that the emotional benefits of WM-T depending more on improved updating and shifting, and that of EWM-T depending more on improved inhibition ability. The advantage to add emotional distractions into general working memory training for targeting rumination related anxiety has been discussed., (Copyright © 2020. Published by Elsevier Inc.)

Attention bias modification (ABM) is a potential intervention in relieving social anxiety symptoms, while its underlying neural mechanisms are not yet understood. The current study included 63 college students with social anxiety. Participants were assigned to the attention modification program (AMP, n = 20), the attention control condition (ACC, n = 20) and the passive waiting group (PW, n = 23). Questionnaires and the emotional Stroop task with EEG recordings were used to assess whether and how the 4-week ABM period affected emotional symptoms and specific emotional processing. Results showed that the two training groups (AMP and ACC) produced comparable emotional improvements and both showed a decrease in negative bias compared with the PW group. The ERP results indicated that despite no significant ERP changes in the PW group, the ACC group exhibited a greater N1, whereas the AMP group exhibited a reduced VPP at the post-test stage compared to the pre-test stage. Besides, both training groups showed a similar late positive potential (LPP) reduction. Notably, the reduction in LPP was positively correlated with behavioral and symptom improvement. Thus, manipulations unique to ABM (face-target contingency) primarily modulate the early attention distribution of material-related stimuli. However, the clinical benefits of attention training may be due to later cognitive-affective mechanisms., (© The Author(s) 2020. Published by Oxford University Press.)

Triple-negative breast cancer (TNBC), which lacks estrogen receptor α (ERα), progesterone receptor, and human epidermal growth factor receptor 2 (HER2) expression, is closely related to basal-like breast cancer. Previously, we and others report that cyclin E/cyclin-dependent kinase 2 (CDK2) phosphorylates enhancer of zeste homolog 2 (EZH2) at T416 (pT416-EZH2). Here, we show that transgenic expression of phospho-mimicking EZH2 mutant EZH2 T416D in mammary glands leads to tumors with TNBC phenotype. Coexpression of EZH2 T416D in mammary epithelia of HER2/Neu transgenic mice reprograms HER2-driven luminal tumors into basal-like tumors. Pharmacological inhibition of CDK2 or EZH2 allows re-expression of ERα and converts TNBC to luminal ERα-positive, rendering TNBC cells targetable by tamoxifen. Furthermore, the combination of either CDK2 or EZH2 inhibitor with tamoxifen effectively suppresses tumor growth and markedly improves the survival of the mice bearing TNBC tumors, suggesting that the mechanism-based combination therapy may be an alternative approach to treat TNBC.

Objective: Previous studies have employed self-report measures to investigate emotion regulation (ER) strategy biases in individuals with anxiety. We investigated the neural signatures underlying ER strategy biases., Method: Twenty individuals with high trait anxiety (HTA) and twenty individuals with low trait anxiety (LTA) completed both the Emotion Regulation Questionnaire and ER tasks. During the tasks, participants were required to passively view, reappraise, and suppress expression while viewing negative images. Event-related potential (ERP) indexes: P2 (an early positive component of the latency around 200ms) and late positive potential (LPP) were adopted to examine the reliability of ER strategy bias in early and later stages during ER procedure., Results: Results of the questionnaire indicated that trait anxiety level was positively correlated with habitual suppression use. ERP results revealed that, in LTA individuals, P2 amplitudes were reduced during both reappraisal and suppression strategy use. HTA individuals, however, showed no significant differences in P2 amplitudes between passive-view and regulation conditions. Furthermore, during the reappraisal block, a reduction of LPP was only observed in the LTA group, while HTA individuals showed a relatively more pronounced reduction in LPP during the application of the suppression strategy., Conclusion: Individuals in the HTA group do not demonstrate the application of effectively strategies during the initial stages of ER process, perhaps due to the presence of an automatic negative bias. While during the later stages of ER, HTA individuals demonstrated successful use of suppression but were deficient in voluntarily applying reappraisal techniques. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Objective: CT120B gene is a splicing variant of CT120A, which deletes 96 nucleotides and leads to an in-frame loss of 32 amino acids between the codon 136 and 167 as compared with CT120A. This study was undertaken to assess the effects of CT120B expression on lung cancer cell growth and to explore the gene expression profiles., Methods: CT120B cDNA was transfected into the human lung adenocarcinoma SPC-A-1 cells, and stable cell lines overexpressing CT120B were established. CCK-8 assay and tumorigenecity in a xenograft model were performed to analyze cell proliferation in vitro and in vivo. The differential gene expression induced by overexpressed CT120B was investigated using Atlas cDNA expression array. Flow cytometry was performed to analyze cell cycle and cell apoptosis., Results: Overexpression of CT120B in SPC-A-1 cells resulted in a reduced cell growth rate in vitro, and decrease of the tumorigenicity in nude mice. A total of 38 genes were identified as differential expressions with more than a 2.0-fold change by Atlas cDNA expression array analysis, including downregulated cyclin E1, cdk 2, c-kit, CXCR4 and upregulated caspase 8 gene. Overexpression of CT120B also induced G1 phase arrest, but had no effect on cell apoptosis., Conclusion: The G1 cell cycle arrest, but not apoptosis, underlay the growth inhibitory activities of CT120B. The down-regulation of c-kit and CXCR4 expression might also contribute to the suppressive effects on cell growth of CT120B.

The expression product of ct120a, a novel gene isolated from human chromosome 17p13.3 in our laboratory, was predicted to have seven transmembrane domains and could cause malignant transformation of mouse NIH3T3 cells. There existed an mRNA splicing variant of ct120a, namely ct120b, which had a 96-nucleotide deletion and produced an in-frame loss of 32 amino acids from codon 136 to codon 167 of CT120A. The CT120B protein was predicted to have six transmembrane domains. In this study, we observed that the green fluorescent protein-tagged CT120B was localized on plasma membrane and in cytoplasm in SPC-A-1 cells. The expression of CT120B/A in normal lung tissue and in lung cancer cells was also examined. Results showed that the stable CT120B overexpression in SPC-A-1 cells resulted in a reduction of cell growth rate, and inhibited tumorigenecity and anchorage-independent growth in nude mice. The functions of CT120A and CT120B for cell growth appeared antagonistic. We suggested that the delayed G1/S phase transition might contribute to the inhibitory activities of CT120B on cell growth and that the deleted 32 amino acids missing in CT120B might be essential for the oncogenetic activities of CT120A.

Objective: To validate our obtained outcomes and clarify the relationship between CT120A, a novel human plasma membrane-associated gene, and proliferation of lung cancer cells., Methods: A vector-based small hairpin RNA (shRNA) was transfected into the human lung adenocarcinoma SPC-A-1 cells to specifically target CT120A cDNA. RT-PCR and Western blotting were used to analyze the CT120A expression. The cell proliferation rate was analyzed by BrdU-TdR incorporation assay, the ability of cells to grow in soft agarose and the tumorigenicity in nude mice were measured. Flow cytometry was performed to analyze cell apoptosis., Results: When compared with the scrambled control cell line, CT120A transcripts were reduced by 70% and 50% in two shRNA-H stable transfectants, H2 and H3 clones, respectively. The protein of CT120A was reduced by about 80% in both the H2 and H3 clones. By BrdU incorporation assay, up to the 6th day a dramatic decrease in the cell growth rate (30% to 40%) was observed in the shRNA-H2 and shRNA-H3 cell lines. The colony formation rate in soft agarose of the two cell lines was about one half that of the control cells. In addition, a remarkable reduction of tumorigenicity of the two cell lines was observed as compared with that of the control. The suppression of CT120A expression also sensitized cells to ultraviolet-induced apoptosis., Conclusion: Down-regulation of CT120A by RNA interference suppresses lung cancer cell growth. The successful knockdown of CT120A expression by RNA interference implicates that CT120A may be a new candidate of drug target for treatment of lung cancers.

Objective: To develop a DNA vector-based RNA interference (RNAi) technology that inhibits the activity of the telomerase of the hepatocellular carcinoma cell line HCCLM3 in order to suppress the proliferation of the cells., Methods: Hepatocellular carcinoma cells of the line HCCLM3 were cultured. mRNA interfering double-stranded DNA vector PSG-AS targeting the mRNA of human telomerase reverse transcriptase (hTERT) and the control vector PSG-CTR were constructed respectively, and then were transfected into the HCCLM3 cells. The expression of hTERT of the transfected cells was determined by Western blotting and the activity of telomerase was determined by telomeric repeat amplification-ELISA (TRAP-ELISA). Flow cytometry was used to detect the apoptosis of transfected cells and MTS method was used to measure the growth curve of the cells so as to observe the effect of the PSG-AS on the proliferation of HCCLM3 cell., Results: TRAP-ELISA showed that the inhibition rate of PSG-AS on the telomerase activity was 76%. The apoptotic rate of the PSG-AS group was significantly higher than that of the PSG-CTR group (t = 11.48, P < 0.001). Western blotting showed a remarkable inhibition of hTERT protein in the PSG-AS group., Conclusion: Capable of suppressing the hTERT expression and the activity of telomerase, RNA interfering technology can be applied to treatment of tumors.

Ribonuclease inhibitor (RI) is an acidic cytosolic glycoprotein with molecular weight of about 50 kDa, which contains 32 cysteine residues. It is possibly that RI may have antioxidant effect by thiol-disulfide exchange reaction. We studied the effects of RI over-expression on the rat glial cell line C6 injured with H2O2. The transfected C6 cells with RI cDNA (C6') had higher viability, less LDH leakage and MDA contents, but more GSH contents compare that in the control C6 cells. In transfected C6 cells, the activities of CAT and GST were higher than that in the control C6 cells. Without H202 stress, the activities of CAT and GST in the C6' cells were 1.73 and 3.62 times that in the control C6 cells, respectively; With 1.00 mmol/L H2O2 stress, the activities of CATand GSTin the C6' cells were 3.38 and 2.11 times that in the C6 cells, respectively. These results suggest that the over-expression RI has antioxidant activity and it is able to protect cells from per-oxidative injuries. Moreover, we investigated whether RI has a protective role against mouse hepatic damage in vivo. The mice pretreated with different doses of human RI were injected by CC14. The results show that the SOD activities of therapy groups were significantly higher than that of the control group (p < 0.01), while the contents of MOD and activities of ALT and AST in blood were remarkably lower than that of the control group (p < 0.01). Pathological examination shows that the degree of damage was alleviated with RI therapy. These results suggest that RI has the protective role against mouse hepatic damage induced by CC14. The anti-oxidative effects of RI may play an important role in cell protection from per-oxidative injuries.