Geological reservoir and resource potential(10^(13)m^(3))of gas hydrates in the South China Sea

A detailed understanding of the distribution and potential of natural gas hydrate(NGHs)resources is crucial to fostering the industrialization of those resources in the South China Sea,where NGHs are abundant.In this study,this study analyzed the applicability of resource evaluation methods,including the volumetric,genesis,and analogy methods,and estimated NGHs resource potential in the South China Sea by using scientific resource evaluation methods based on the factors controlling the geological accumulation and the reservoir characteristics of NGHs.Furthermore,this study compared the evaluation results of NGHs resource evaluations in representative worldwise sea areas via rational analysis.The results of this study are as follows:(1)The gas hydrate accumulation in the South China Sea is characterized by multiple sources of gas supply,multi-channel migration,and extensive accumulation,which are significantly different from those of oil and gas and other unconventional resources.(2)The evaluation of gas hydrate resources in the South China Sea is a highly targeted,stratified,and multidisciplinary evaluation of geological resources under the framework of a multi-type gas hydrate resource evaluation system and focuses on the comprehensive utilization of multi-source heterogeneous data.(3)Global NGHs resources is n×10^(15)m^(3),while the NGHs resources in the South China Sea are estimated to be 10^(13)m^(3),which is comparable to the abundance of typical marine NGHs deposits in other parts of the world.In the South China Sea,the NGHs resources have a broad prospect and provide a substantial resource base for production tests and industrialization of NGHs.

Gas-Water Production of a Continental Tight-Sandstone Gas Reservoir under Different Fracturing Conditions

A numerical model of hydraulic fracture propagation is introduced for a representative reservoir(Yuanba continental tight sandstone gas reservoir in Northeast Sichuan).Different parameters are considered,i.e.,the interlayer stress difference,the fracturing discharge rate and the fracturing fluid viscosity.The results show that these factors affect the gas and water production by influencing the fracture size.The interlayer stress difference can effectively control the fracture height.The greater the stress difference,the smaller the dimensionless reconstruction volume of the reservoir,while the flowback rate and gas production are lower.A large displacement fracturing construction increases the fracture-forming efficiency and expands the fracture size.The larger the displacement of fracturing construction,the larger the dimensionless reconstruction volume of the reservoir,and the higher the fracture-forming efficiency of fracturing fluid,the flowback rate,and the gas production.Low viscosity fracturing fluid is suitable for long fractures,while high viscosity fracturing fluid is suitable for wide fractures.With an increase in the fracturing fluid viscosity,the dimensionless reconstruction volume and flowback rate of the reservoir display a non-monotonic behavior,however,their changes are relatively small.

Hydrocarbon gas huff-n-puff optimization of multiple horizontal wells with complex fracture networks in the M unconventional reservoir

The oil production of the multi-fractured horizontal wells(MFHWs) declines quickly in unconventional oil reservoirs due to the fast depletion of natural energy. Gas injection has been acknowledged as an effective method to improve oil recovery factor from unconventional oil reservoirs. Hydrocarbon gas huff-n-puff becomes preferable when the CO_(2) source is limited. However, the impact of complex fracture networks and well interference on the EOR performance of multiple MFHWs is still unclear. The optimal gas huff-n-puff parameters are significant for enhancing oil recovery. This work aims to optimize the hydrocarbon gas injection and production parameters for multiple MFHWs with complex fracture networks in unconventional oil reservoirs. Firstly, the numerical model based on unstructured grids is developed to characterize the complex fracture networks and capture the dynamic fracture features.Secondly, the PVT phase behavior simulation was carried out to provide the fluid model for numerical simulation. Thirdly, the optimal parameters for hydrocarbon gas huff-n-puff were obtained. Finally, the dominant factors of hydrocarbon gas huff-n-puff under complex fracture networks are obtained by fuzzy mathematical method. Results reveal that the current pressure of hydrocarbon gas injection can achieve miscible displacement. The optimal injection and production parameters are obtained by single-factor analysis to analyze the effect of individual parameter. Gas injection time is the dominant factor of hydrocarbon gas huff-n-puff in unconventional oil reservoirs with complex fracture networks. This work can offer engineers guidance for hydrocarbon gas huff-n-puff of multiple MFHWs considering the complex fracture networks.

Fine quantitative characterization of high-H2S gas reservoirs under the influence of liquid sulfur deposition and adsorption

In order to clarify the influence of liquid sulfur deposition and adsorption to high-H2S gas reservoirs,three types of natural cores with typical carbonate pore structures were selected for high-temperature and high-pressure core displacement experiments.Fine quantitative characterization of the cores in three steady states(original,after sulfur injection,and after gas flooding)was carried out using the nuclear magnetic resonance(NMR)transverse relaxation time spectrum and imaging,X-ray computer tomography(CT)of full-diameter cores,basic physical property testing,and field emission scanning electron microscopy imaging.The loss of pore volume caused by sulfur deposition and adsorption mainly comes from the medium and large pores with sizes bigger than 1000μm.Liquid sulfur has a stronger adsorption and deposition ability in smaller pore spaces,and causes greater damage to reservoirs with poor original pore structures.The pore structure of the three types of carbonate reservoirs shows multiple fractal characteristics.The worse the pore structure,the greater the change of internal pore distribution caused by liquid sulfur deposition and adsorption,and the stronger the heterogeneity.Liquid sulfur deposition and adsorption change the pore size distribution,pore connectivity,and heterogeneity of the rock,which further changes the physical properties of the reservoir.After sulfur injection and gas flooding,the permeability of TypeⅠreservoirs with good physical properties decreased by 16%,and that of TypesⅡandⅢreservoirs with poor physical properties decreased by 90%or more,suggesting an extremely high damage.This indicates that the worse the initial physical properties,the greater the damage of liquid sulfur deposition and adsorption.Liquid sulfur is adsorbed and deposited in different types of pore space in the forms of flocculence,cobweb,or retinitis,causing different changes in the pore structure and physical property of the reservoir.

Development and technology status of energy storage in depleted gas reservoirs

Utilizing energy storage in depleted oil and gas reservoirs can improve productivity while reducing power costs and is one of the best ways to achieve synergistic development of"Carbon Peak–Carbon Neutral"and"Underground Resource Utiliza-tion".Starting from the development of Compressed Air Energy Storage(CAES)technology,the site selection of CAES in depleted gas and oil reservoirs,the evolution mechanism of reservoir dynamic sealing,and the high-flow CAES and injection technology are summarized.It focuses on analyzing the characteristics,key equipment,reservoir construction,application scenarios and cost analysis of CAES projects,and sorting out the technical key points and existing difficulties.The devel-opment trend of CAES technology is proposed,and the future development path is scrutinized to provide reference for the research of CAES projects in depleted oil and gas reservoirs.

Horizontal well spacing optimization and gas injection simulation for the ultra-low-permeability Yongjin reservoir

Optimal spacing for vertical wells can be effectively predicted with several published methods,but methods suitable for assessing the proper horizontal well spacing are rare.This work proposes a method for calculating the optimal horizontal well spacing for an ultra-low permeability reservoir e the Yongjin reservoir in the Juggar Basin,northwestern China.The result shows that a spacing of 640m is the most economical for the development of the reservoir.To better develop the reservoir,simulation approaches are used and a new model is built based on the calculated well spacing.Since the reservoir has an ultralow permeability,gas injection is regarded as the preferred enhanced oil recovery(EOR)method.Injection of different gases including carbon dioxide,methane,nitrogen and mixed gas are modelled.The results show that carbon dioxide injection is the most efficient and economical for the development of the reservoir.However,if the reservoir produces enough methane,reinjecting methane is even better than injecting carbon dioxide.

Optimization of Gas-Flooding Fracturing Development in Ultra-Low Permeability Reservoirs

Ultra-low permeability reservoirs are characterized by small pore throats and poor physical properties, which areat the root of well-known problems related to injection and production. In this study, a gas injection floodingapproach is analyzed in the framework of numerical simulations. In particular, the sequence and timing of fracturechanneling and the related impact on production are considered for horizontal wells with different fracturemorphologies. Useful data and information are provided about the regulation of gas channeling and possible strategiesto delay gas channeling and optimize the gas injection volume and fracture parameters. It is shown that inorder to mitigate gas channeling and ensure high production, fracture length on the sides can be controlled andlonger fractures can be created in the middle by which full gas flooding is obtained at the fracture location in themiddle of the horizontal well. A Differential Evolution (DE) algorithm is provided by which the gas injectionvolume and the fracture parameters of gas injection flooding can be optimized. It is shown that an improvedoil recovery factor as high as 6% can be obtained.

A new model simulating the development of gas condensate reservoirs

A new simulation model for the development of gas condensate reservoirs is introduced based on the influence that phase change,non-Darcy flow,and capillary pressure have on the production of gas condensates.The model predicts well performance,including bottom-hole pressure,oil/gas production rate,oil/gas recovery,gaseoil ratio,and the change in produced fluid composition.It also calculates dynamic characters,such as the change of pressure field and oil/gas saturation field during the development of gas condensate reservoirs.The model is applicable to different boundary conditions(both constant-pressure and sealed boundary)and different production modes(both constant-pressure and constant-volume production modes).Model validation attempted using numerical simulation results for sealed boundary conditions with constant-pressure production mode has shown a relatively good match,proving its validity.For constant-pressure boundary conditions with constant-volume production mode,four stages are defined according to the dynamic behavior of production performance in the development of gas condensate reservoirs.

Evaluation of Well Spacing for Primary Development of Fractured Horizontal Wells in Tight Sandstone Gas Reservoirs

Methods for horizontal well spacing calculation in tight gas reservoirs are still adversely affected by the complexity of related control factors,such as strong reservoir heterogeneity and seepage mechanisms.In this study,the stress sensitivity and threshold pressure gradient of various types of reservoirs are quantitatively evaluated through reservoir seepage experiments.On the basis of these experiments,a numerical simulation model(based on the special seepage mechanism)and an inverse dynamic reserve algorithm(with different equivalent drainage areas)were developed.The well spacing ranges of Classes I,II,and III wells in the Q gas field are determined to be 802–1,000,600–662,and 285–400 m,respectively,with their average ranges as 901,631,and 342.5 m,respectively.By considering both the pairs of parallel well groups and series well groups as examples,the reliability of the calculation results is verified.It is shown that the combination of the two models can reduce errors and provide accurate results.

Microscopic experiment on efficient construction of underground gas storages converted from water-invaded gas reservoirs

Based on the microfluidic technology,a microscopic visualization model was used to simulate the gas injection process in the initial construction stage and the bottom water invasion/gas injection process in the cyclical injection-production stage of the underground gas storage(UGS)rebuilt from water-invaded gas reservoirs.Through analysis of the gas-liquid contact stabilization mechanism,flow and occurrence,the optimal control method for lifecycle efficient operation of UGS was explored.The results show that in the initial construction stage of UGS,the action of gravity should be fully utilized by regulating the gas injection rate,so as to ensure the macroscopically stable migration of the gas-liquid contact,and greatly improve the gas sweeping capacity,providing a large pore space for gas storage in the subsequent cyclical injection-production stage.In the cyclical injection-production stage of UGS,a constant gas storage and production rate leads to a low pore space utilization.Gradually increasing the gas storage and production rate,that is,transitioning from small volume to large volume,can continuously break the hydraulic equilibrium of the remaining fluid in the porous media,which then expands the pore space and flow channels.This is conducive to the expansion of UGS capacity and efficiency for purpose of peak shaving and supply guarantee.

Rate transient analysis methods for water-producing gas wells in tight reservoirs with mobile water

Tight gas reservoirs with mobile water exhibit multi-phase flow and high stress sensitivity.Accurately analyzing the reservoir and well parameters using conventional single-phase rate transient analysis methods proves challenging.This study introduces novel rate transient analysis methods incorporating evaluation processes based on the conventional flowing material balance method and the Blasingame type-curve method to examine fractured gas wells producing water.By positing a gas-water two-phase equivalent homogenous phase that considers characteristics of mobile water,gas,and high stress sensitivity,the conventional single-phase rate transient analysis methods can be applied by integrating the phase's characteristics and defining the phase's normalized parameters and material balance pseudotime.The rate transient analysis methods based on the equivalent homogenous phase can be used to quantitatively assess the parameters of wells and gas reservoirs,such as original gas-in-place,fracture half-length,reservoir permeability,and well drainage radius.This facilitates the analysis of production dynamics of fractured wells and well-controlled areas,subsequently aiding in locating residual gas and guiding the configuration of well patterns.The specific evaluation processes are detailed.Additionally,a numerical simulation mechanism model was constructed to verify the reliability of the developed methods.The methods introduced have been successfully implemented in field water-producing gas wells within tight gas reservoirs containing mobile water.

Numerical study on gas production via a horizontal well from hydrate reservoirs with different slope angles in the South China Sea

It is important to study the effect of hydrate production on the physical and mechanical properties of low-permeability clayey–silty reservoirs for the largescale exploitation of hydrate reservoirs in the South China Sea.In this study,a multiphysical-field coupling model,combined with actual exploration drilling data and the mechanical experimental data of hydrate cores in the laboratory,was established to investigate the physical and mechanical properties of low-permeability reservoirs with different slope angles during 5-year hydrate production by the depressurization method via a horizontal well.The result shows that the permeability of reservoirs severely affects gas production rate,and the maximum gas production amount of a 20-m-long horizontal well can reach186.8 m3/day during the 5-year hydrate production.Reservoirs with smaller slope angles show higher gas production rates.The depressurization propagation and hydrate dissociation mainly develop along the direction parallel to the slope.Besides,the mean effective stress of reservoirs is concentrated in the near-wellbore area with the on-going hydrate production,and gradually decreases with the increase of the slope angle.Different from the effective stress distribution law,the total reservoir settlement amount first decreases and then increases with the increase of the slope angle.The maximum settlement of reservoirs with a 0°slope angle is up to 3.4 m,and the displacement in the near-wellbore area is as high as2.2 m after 5 years of hydrate production.It is concluded that the pore pressure drop region of low-permeability reservoirs in the South China Sea is limited,and various slope angles further lead to differences in effective stress and strain of reservoirs during hydrate production,resulting in severe uneven settlement of reservoirs.

Effects of acid-rock reaction on physical properties during CO_(2)-rich industrial waste gas(CO_(2)-rich IWG)injection in shale reservoirs

"Carbon peaking and carbon neutrality"is an essential national strategy,and the geological storage and utilization of CO_(2)is a hot issue today.However,due to the scarcity of pure CO_(2)gas sources in China and the high cost of CO_(2)capture,CO_(2)-rich industrial waste gas(CO_(2)-rich IWG)is gradually emerging into the public's gaze.CO_(2)has good adsorption properties on shale surfaces,but acidic gases can react with shale,so the mechanism of the CO_(2)-rich IWG-water-shale reaction and the change in reservoir properties will determine the stability of geological storage.Therefore,based on the mineral composition of the Longmaxi Formation shale,this study constructs a thermodynamic equilibrium model of water-rock reactions and simulates the regularity of reactions between CO_(2)-rich IWG and shale minerals.The results indicate that CO_(2)consumed 12%after reaction,and impurity gases in the CO_(2)-rich IWG can be dissolved entirely,thus demonstrating the feasibility of treating IWG through water-rock reactions.Since IWG inhibits the dissolution of CO_(2),the optimal composition of CO_(2)-rich IWG is 95%CO_(2)and 5%IWG when CO_(2)geological storage is the main goal.In contrast,when the main goal is the geological storage of total CO_(2)-rich IWG or impurity gas,the optimal CO_(2)-rich IWG composition is 50%CO_(2)and 50%IWG.In the CO_(2)-rich IWG-water-shale reaction,temperature has less influence on the water-rock reaction,while pressure is the most important parameter.SO2 has the greatest impact on water-rock reaction in gas.For minerals,clay minerals such as illite and montmorillonite had a significant effect on water-rock reaction.The overall reaction is dominated by precipitation and the volume of the rock skeleton has increased by 0.74 cm3,resulting in a decrease in shale porosity,which enhances the stability of CO_(2)geological storage to some extent.During the reaction between CO_(2)-rich IWG-water-shale at simulated temperatures and pressures,precipitation is the main reaction,and shale porosity decreases.However,as the reservoir water content increases,the reaction will first dissolve and then precipitate before dissolving again.When the water content is less than 0.0005 kg or greater than 0.4 kg,it will lead to an increase in reservoir porosity,which ultimately reduces the long-term geological storage stability of CO_(2)-rich IWG.

Ordovician reservoirs in Fuxian area:Gas accumulation patterns and their implications for the exploration of lower Paleozoic carbonates in the southern Ordos Basin

In recent years,the Fuxian area in the southeastern Ordos Basin has undergone significant exploration,with industrial gas flow tested in wells drilled into the Ordovician marine carbonates.Despite this,the gas accumulation patterns of this area are not fully understood,posing challenges for further exploration.Our analysis of geological conditions indicates that the Ordovician Majiagou Formation in this area hosts two gas plays:one found in weathering crusts and the other found in interior of the formation.We investigated various typical gas reservoirs in the area,focusing on differentiating the geological conditions and factors controlling gas accumulation in the weathering-crust and interior gas reservoirs.The results suggest three primary gas accumulation patterns in the Majiagou Formation in the Fuxian area:(1)upper gas accumulation in weathering crusts,present in the high parts of landforms such as residual paleo-hills or buried paleo-platform(Pattern I);(2)the stereoscopic pattern with gas accumulation in both weathering crusts and strata interior,arising in high parts of landforms such as residual paleo-hills or buried paleo-platforms(Pattern II);(3)lower gas accumulation in strata interior,occurring in the upper reaches and on both sides of paleo-trenches(Pattern III).This study will serve as a geological basis for future exploration deployment in the Fuxian area.

基于GAS算法的卵砾石粒径自动识别应用研究

粒径和级配是表征床面组成的重要指标,基于GAS粒径自动识别技术可自动识别粗粒床面粒径并生成级配曲线,能够大幅提高现场采样和分析的效率。为了验证GAS的分割效果,采用GAS提供的默认参数进行分割,同时应用ImageJ软件手动分割进行验证。结果表明:GAS级配曲线的相对误差为5.7%,相关系数为0.992。另外,采用单参数和多参数敏感性分析法来标准化参数调整方案,gre、can1和can2对GAS提取的级配曲线和特征粒径有显著影响,其中gre起主导作用,而can1和can2控制着砾石边界的检测完整性。

Geochemistry and origins of hydrogen-containing natural gases in deep Songliao Basin,China:Insights from continental scientific drilling

The different reservoirs in deep Songliao Basin have non-homogeneous lithologies and include multiple layers with a high content of hydrogen gas.The gas composition and stable isotope characteristics vary significantly,but the origin analysis of different gas types has previously been weak.Based on the geochemical parameters of gas samples from different depths and the analysis of geological settings,this research covers the diverse origins of natural gas in different strata.The gas components are mainly methane with a small amount of C_(2+),and non-hydrocarbon gases,including nitrogen(N_(2)),hydrogen(H_(2)),carbon dioxide(CO_(2)),and helium(He).At greater depth,the carbon isotope of methane becomes heavier,and the hydrogen isotope points to a lacustrine sedimentary environment.With increasing depth,the origins of N_(2)and CO_(2)change gradually from a mixture of organic and inorganic to inorganic.The origins of hydrogen gas are complex and include organic sources,water radiolysis,water-rock(Fe^(2+)-containing minerals)reactions,and mantle-derived.The shales of Denglouku and Shahezi Formations,as source rocks,provide the premise for generation and occurrence of organic gas.Furthermore,the deep faults and fluid activities in Basement Formation control the generation and migration of mantle-derived gas.The discovery of a high content of H_(2)in study area not only reveals the organic and inorganic association of natural-gas generation,but also provides a scientific basis for the exploration of deep hydrogen-rich gas.

Repositioning fertilizer manufacturing subsidies for improving food security and reducing greenhouse gas emissions in China

China removed fertilizer manufacturing subsidies from 2015 to 2018 to bolster market-oriented reforms and foster environmentally sustainable practices.However,the impact of this policy reform on food security and the environment remains inadequately evaluated.Moreover,although green and low-carbon technologies offer environmental advantages,their widespread adoption is hindered by prohibitively high costs.This study analyzes the impact of removing fertilizer manufacturing subsidies and explores the potential feasibility of redirecting fertilizer manufacturing subsidies to invest in the diffusion of these technologies.Utilizing the China Agricultural University Agri-food Systems model,we analyzed the potential for achieving mutually beneficial outcomes regarding food security and environmental sustainability.The findings indicate that removing fertilizer manufacturing subsidies has reduced greenhouse gas(GHG)emissions from agricultural activities by 3.88 million metric tons,with minimal impact on food production.Redirecting fertilizer manufacturing subsidies to invest in green and low-carbon technologies,including slow and controlled-release fertilizer,organic-inorganic compound fertilizers,and machine deep placement of fertilizer,emerges as a strategy to concurrently curtail GHG emissions,ensure food security,and secure robust economic returns.Finally,we propose a comprehensive set of government interventions,including subsidies,field guidance,and improved extension systems,to promote the widespread adoption of these technologies.

Geochemical characteristics and exploration significance of ultra-deep Sinian oil and gas from Well Tashen 5,Tarim Basin,NW China

The Well Tashen 5(TS5),drilled and completed at a vertical depth of 9017 m in the Tabei Uplift of the Tarim Basin,NW China,is the deepest well in Asia.It has been producing both oil and gas from the Sinian at a depth of 8780e8840 m,also the deepest in Asia in terms of oil discovery.In this paper,the geochemical characteristics of Sinian oil and gas from the well were investigated and compared with those of Cambrian oil and gas discovered in the same basin.The oil samples,with Pr/Ph ratio of 0.78 and a whole oil carbon isotopic value of31.6‰,have geochemical characteristics similar to those of Ordovician oils from the No.1 fault in the North Shuntuoguole area(also named Shunbei area)and the Middle Cambrian oil from wells Zhongshen 1(ZS1)and Zhongshen 5(ZS5)of Tazhong Uplift.The maturity of light hydrocarbons,diamondoids and aromatic fractions all suggest an approximate maturity of 1.5%e1.7%Ro for the samples.The(4-+3-)methyldiamantane concentration of the samples is 113.5 mg/g,indicating intense cracking with a cracking degree of about 80%,which is consistent with the high bottom hole temperature(179℃).The Sinian gas samples are dry with a dryness coefficient of 0.97.The gas is a mixture of kerogen-cracking gas and oil-cracking gas and has Ro values ranging between 1.5%and 1.7%,and methane carbon isotopic values of41.6‰.Based on the equivalent vitrinite reflectance(R_(eqv)=1.51%e1.61%)and the thermal evolution of source rocks from the Cambrian Yu'ertusi Formation of the same well,it is proposed that the Sinian oil and gas be mainly sourced from the Cambrian Yu'ertusi Formation during the Himalayan period but probably also be joined by hydrocarbon of higher maturity that migrated from other source rocks in deeper formations.The discovery of Sinian oil and gas from Well TS5 suggests that the ancient ultra-deep strata in the northern Tarim Basin have the potential for finding volatile oil or condensate reservoirs.

GAS6-AS1调节miR-370-3p/SPATA2轴对卵巢癌细胞增殖、迁移、侵袭、凋亡和EMT的影响

目的:探讨长链非编码RNA GAS6反义RNA1(long non-coding RNA GAS6 antisense RNA1, lncRNA GAS6-AS1)调节miR-370-3p/精子发生相关蛋白2 (spermatogenesis-associated protein 2, SPATA2)轴对卵巢癌细胞增殖、迁移、侵袭、凋亡和上皮间质转化(epithelial mesenchymal transformation, EMT)的影响。方法:qRT-PCR、Western blot分别检测癌旁组织、卵巢癌组织、人正常卵巢上皮细胞IOSE80及卵巢癌细胞系HO-8910、SKOV3、A2780中GAS6-AS1、miR-370-3p及SPATA2蛋白表达。将SKOV3细胞分为:对照组(NC组)、 si-NC组、si-GAS6-AS1组、mimic NC组、miR-370-3p mimic组、si-GAS6-AS1+inhibitor NC组、si-GAS6-AS1+miR-370-3p inhibitor组,qRT-PCR检测细胞中GAS6-AS1、miR-370-3p表达;CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;Western blot检测SPATA2、细胞周期素D1(CyclinD1)、Bcl-2相关X蛋白(Bcl-2-associated X,Bax)、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、神经钙黏蛋白(N-cadherin)表达;双荧光素酶报告基因实验检测GAS6-AS1与miR-370-3p、 miR-370-3p与SPATA2的关系。结果:在卵巢癌组织和细胞中GAS6-AS1、SPATA2蛋白高表达,miR-370-3p低表达,且在SKOV3细胞中GAS6-AS1、SPATA2蛋白表达量最高,miR-370-3p表达水平最低,因此,选择SKOV3细胞为后续研究对象。与NC组、si-NC组比较,si-GAS6-AS1组GAS6-AS1、OD450值(24 h、48 h、72 h)、划痕愈合率、侵袭细胞数、SPATA2、CyclinD1、Vimentin、N-cadherin蛋白表达降低,miR-370-3p表达、细胞凋亡率、Bax、E-cadherin蛋白表达升高(P<0.05);与NC组、mimic NC组比较,miR-370-3p mimic组OD450值(24 h、48 h、72 h)、划痕愈合率、侵袭细胞数、SPATA2、CyclinD1、Vimentin、N-cadherin蛋白表达降低,miR-370-3p表达、细胞凋亡率、Bax、E-cadherin蛋白表达升高(P<0.05);miR-370-3p inhibitor减弱了沉默GAS6-AS1对SKOV3细胞增殖、迁移、侵袭及EMT的抑制及对细胞凋亡的促进作用。GAS6-AS1与miR-370-3p、miR-370-3p与SPATA2存在靶向调控关系。结论:沉默GAS6-AS1通过上调miR-370-3p来抑制SPATA2表达,从而抑制SKOV3细胞增殖、迁移、侵袭及EMT,并促进细胞凋亡。

LncRNA GAS5-AS1靶向miR-106a-5p调控结直肠癌细胞增殖、迁移和侵袭

背景长链非编码RNA(long non-coding RNA,LncRNA)生长停滞特异性转录本5反义RNA(growth arrest specific transcript 5 antisense RNA,GAS5-AS1)被发现在多种癌症中扮演肿瘤抑制因子的作用.但是其在结直肠癌(colorectal cancer,CRC)进展中的作用和机制尚不清楚.目的探讨LncRNA GAS5-AS1靶向miR-106a-5p在CRC进展中的作用.方法实时荧光定量聚合酶链反应分析LncRNA GAS5-AS1和miR-106a-5p在43对CRC组织和癌旁组织样本中的表达.分别转染pcDNA、pcDNA-LncRNA GAS5-AS1、anti-miR-NC、anti-miR-106a-5p、si-NC、si-LncRNA GAS5-AS1、pcDNA-LncRNA GAS5-AS1+miR-NC、pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics至HCT8细胞.细胞计数试剂盒-8法检测细胞活力;Transwell小室法检测迁移和侵袭细胞数.通过双荧光素酶报告实验确定LncRNA GAS5-AS1和miR-106a-5p的靶向关系.结果与癌旁组织比较,CRC组织中LncRNA GAS5-AS1表达降低(P<0.05),miR-106a-5p表达升高(P<0.05).与转染pcDNA比较,转染pcDNA-LncRNA GAS5-AS1后HCT8细胞活力、迁移、侵袭、miR-106a-5p表达显著降低(P<0.05).与转染anti-miR-NC比较,转染anti-miR-106a-5p后HCT8细胞活力、迁移和侵袭细胞数显著降低(P<0.05).与转染si-NC比较,转染si-LncRNA GAS5-AS1后miR-106a-5p表达水平显著升高(P<0.05).与转染miR-NC比较,转染miR-106a-5p能显著降低LncRNA GAS5-AS1-WT载体的荧光素酶活性,表明miR-106a-5p是LncRNA GAS5-AS1的直接靶点.与转染pcDNA-LncRNA GAS5-AS1+miR-NC比较,转染pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics后HCT8细胞活力、迁移和侵袭细胞数显著升高(P<0.05).结论LncRNA GAS5-AS1通过靶向miR-106a-5p来抑制CRC细胞增殖、迁移和侵袭.